Compositions and methods for treating or preventing pneumovirus infection and associated diseases

ABSTRACT

The present invention provides novel crystalline polymorphic forms of MDT-637, in particular, crystalline polymorphic forms with physicochemical properties specifically suited for drug production, amorphous formation, composite form, and methods of preparation thereof. The novel polymorphs described herein are useful for the treatment of respiratory disease, such as disease caused by respiratory syncytial virus.

CROSS REFERENCE TO RELATED APPLICATION

The present application claims priority from U.S. ProvisionalApplication Ser. No. 61/666,258 filed Jun. 29, 2012 the contents ofwhich are incorporated herein in their entirety, by reference.

FIELD OF THE INVENTION

The present invention relates to compounds, compositions and methods forpreventing and treating viral infections, and the diseases associatedtherewith, particularly those viral infections and diseases caused byviruses of the order Paramyxoviridae, including Paramyxovirinae andPneumovirinae subfamilies. More specifically, the present inventionrelates to novel crystalline polymorphic forms of MDT-637, inparticular, crystalline polymorphic forms with physicochemicalproperties advantageous for drug product, amorphous form, compositeform, and methods of preparation thereof.

BACKGROUND OF THE INVENTION

MDT-637 is an active pharmaceutical ingredient (API) (chemical name:phenol,2,2′-[(4-hydroxyphenyl)methylene]bis[4-[[(5-methyl-1H-tetrazol-1-yl)imino]methyl]];alternate name:5,5′-Bis[1-(((5-methyl-1-H-tetrazolyl)imino)methyl)]-2,2′,4″-methylidynetrisphenol; molecular formula C₂₅H₂₂N₁₀O₃) as described by the followingstructure:

MDT-637 exhibits an antiviral therapeutic activity as described in U.S.Pat. No. 6,495,580 which is hereby incorporated by reference. Inaddition, U.S. patent application Ser. No. 10/524,162 and U.S. patentapplication Ser. No. 10/524,313 describe related compounds andcompositions and are also incorporated by reference. MDT-637 isassociated with preventing and treating viral infections, and thediseases associated therewith, particularly those viral infections anddiseases caused by viruses of the order Paramyxoviridae, includingParamyxovirinae and Pneumovirinae subfamilies.

A number of important human diseases are caused by Paramyxoviruses,including mumps, measles, and respiratory syncytial virus (RSV), whichis a major cause of bronchiolitis and pneumonia in infants and childrenboth in the US and worldwide. The mechanism of action of MDT-637 hasbeen elucidated in some detail, and though not wishing to be bound bythe following theory, it is thought that MDT-637 acts by targeting andblocking the viral fusion protein, which is a target for RSV treatments.In addition to being highly potent (40,000 times more potent thanribavirin) MDT-637 also showed that it was effective in reducing RSVviral count both pre- and post-infection.

The family Paramyxoviridae is composed of a diverse group of viruses andis divided into two subfamilies, Paramyxovirinae and Pneumovirinae.

The major human viruses of the Paramyxoviridae family are: measlesvirus, mumps virus, the parainfluenza viruses (types 1, 2, 3, 4a, and4b), and respiratory syncytial virus (RSV). All of the viruses of theParamyxoviridae family are spread through the respiratory route and arehighly contagious. A number of important human diseases are caused byparamyxoviruses. These include mumps, measles, which caused 745,000deaths in 2001 and respiratory syncytial virus (RSV), which is a majorcause of bronchiolitis and pneumonia in infants and children both in theUS and worldwide.

Paramyxoviruses are also responsible for a range of diseases in otheranimal species, for example canine distemper virus (dogs), phocinedistemper virus (seals), cetacean morbillivirus (dolphins and porpoises)Newcastle disease virus (birds), and rinderpest virus (cattle). Someparamyxoviruses such as the henipaviruses are zoonotic pathogens,occurring naturally in an animal host, but also able to infect humans.Also included are certain “unassigned” viruses, such as Atlantic salmonparamyxovirus, Beilong virus, J virus, Pacific salmon paramyxovirus, andTailam virus.

The parainfluenza viruses are the second most common causes ofrespiratory tract disease in infants and children. They can causepneumonia, bronchitis and croup in children and the elderly. Infectionwith parainfluenza viruses typically produce minor upper respiratorytract infections which are characterized by coryza, pharyngitis, lowfever, and bronchitis. Parainfluenza viruses are also the most commoncause of croup, or laryngotracheobronchitis, in children aged 6 monthsto 5 years.

Human RSV, the prototypic member of the pneumovirus group, is the majorpediatric viral respiratory tract pathogen, causing pneumonia andbronchiolitis in infants and young children. According to the USNational Institutes of Health, human RSV infection, the single mostimportant cause of severe respiratory illness in infants and youngchildren and the major cause of infantile bronchiolitis, is the mostfrequent cause of hospitalization of infants and young children inindustrialized countries. In the USA alone, from 85,000 to 144,000infants with RSV infection are hospitalized annually, resulting in20%-25% of pneumonia cases and up to 70% of bronchiolitis cases in thehospital. Global RSV disease burden is estimated at 64 million cases and160,000 deaths every year.

Children who experience RSV infection early in life run a high risk ofsubsequent recurrent wheezing and asthma, especially premature infantsand infants with bronchopulmonary dysplasia, for whom preventive passiveimmunization with anti-RSV monoclonal antibodies such as Palivizumab ishighly recommended. RSV also is a significant problem in the elderly, inpersons with cardiopulmonary diseases and in immunocompromizedindividuals. RSV attack rates in nursing homes in the USA areapproximately 5%-10% per year with a 2%-8% case fatality rate, amountingto approximately 10,000 deaths per year among persons older than 64years of age.

Attempts to develop vaccines for RSV are ongoing, but none have yet beendemonstrated to be safe and efficacious. Vaccine development has beenshadowed by adverse reactions exhibited by the initialformalin-inactivated RSV vaccine introduced in the late 1960s. Immunizedchildren showed an increased incidence of RSV lower respiratory tractdisease and developed abnormally severe illnesses, including death.Chemotherapy with ribavirin[1-beta-D-ribofuranosyl-1H-1,2,4-triazole-3-carboxamide], an antiviralnucleoside which is the only pharmaceutical approved by the U.S. Foodand Drug Administration (FDA) for treatment of RSV disease, isconsidered only for certain RSV patients (e.g., those at high risk forsevere complications or who are seriously ill with this infection).However, its efficacy and value are controversial. Recent studies havereported a failure to demonstrate either clinical or economic benefit topatients of ribavirin treatment. Moreover, ribavirin has certain toxicside-effects and, in order to minimize these, strict administrativeprocedures in a closed environment must be followed.

A human intravenous immune globulin (IVIG) preparation is licensed forprophylactic use in certain patients at high-risk for RSV disease.Administration of this drug requires intravenous infusion of a largevolume over a 2 to 4 hour period in children who have limited venousaccess due to prior intensive therapy, as well as compromisedcardiopulmonary function. Moreover, intravenous infusion necessitatesmonthly hospital visits during the RSV season, which in turn placeschildren at risk of nosocomial infections.

None of the above-described regimens satisfies the need for effectivevaccines or therapeutics for RSV infection. Given the high risk ofoccurrence, along with the high incidence of mortality amongstvulnerable populations (pediatric, immunocompromised, elderly), there isa clear need for novel and effective therapeutic regimens that canalleviate and eliminate the complications associated with pneumovirusinfections.

As described above, there is a wide array of infections associated withParamyxovirinae and Pneumovirinae pathogens, and accordingly, it isevident that there exists a need for improved therapeutic regimens thatcan alleviate and eliminate complications associated with infection.Currently, no such therapeutics are available. In particular, a needexists for new anti-viral agents and treatments for RSV infection thatovercome the shortcomings of existing pharmaceutical preparations.

MDT-637 is recognized as an effective antiviral compound, but therecontinues to be a need for improved compositions having desirabletherapeutic characteristics such as modes of delivery and optimizeddistribution allowing effective and safe dosing. In addition, there is aneed for compositions with beneficial physical and chemical properties,stability and handling characteristics. Furthermore, there is need fornovel and consistently predictable methods of manufacturing, therebyreducing the potential for heterogeneous compositions.

SUMMARY OF THE INVENTION

The present invention provides an improvement over prior art vaccinesand therapeutics by providing novel compositions for the treatment andprevention of infection caused by, or associated with, Paramyxovirinaeand Pneumovirinae infection. In particular, the novel compositions andmethods of the present invention are well suited to therapeuticintervention in infection caused by the major viruses of theParamyxoviridae family including, but not limited to, measles virus,mumps virus, the parainfluenza viruses (types 1, 2, 3, 4a, and 4b), andrespiratory syncytial virus (RSV). The improved compositions and methodsof the present invention satisfy the heretofore unmet need in the artfor therapeutic intervention that enables the alleviation of symptomssuch as rhinitis, otitis media, pneumonia and bronchiolitis. Inaddition, the novel compositions and methods of the present inventionare particularly desirable due to improved stability, improvedsolubility, low dosing levels, ease of handling, dosing andadministration as well as the significant reduction and absence of sideeffects and toxicity. Surprisingly, the compositions described hereindisplay unexpected results with regard to therapeutic efficacy.

The methods and compositions described herein are particularly suitedfor treating Paramyxovirinae and Pneumovirinae infection, however, aswould be evident to one skilled in the art, they may also be utilizedfor additional indications.

The present invention provides new crystal forms of MDT-637 with uniquestructure, hydration or solvation levels as well as novel methods oftheir production.

The present invention provides a novel crystal form, denominated as formof pattern MDT-637 P-3 dihydrate (or simply P-3 dihydrate) which isfound to be particularly advantageous for drug delivery of MDT-637.

In another aspect, the present invention provides a novel crystal formof P-3 ethanolate which plays an important role in purification andproduction of P-3 dihydrate.

In another aspect, the present invention provides a novel crystal formof P-3 monohydrate.

In another aspect, the present invention provides a novel crystal formof P-3 anhydrous.

In another aspect, the present invention provides an additional novelMDT-637 P-2 crystal form, denominated as form of pattern P-2 hydrate (orsimply P-2 hydrate) which is shown to be the most thermodynamicallystable form at ambient conditions.

In another aspect, the present invention provides a novel crystal form,P-2 anhydrous.

In another aspect, the present invention provides a purificationrecrystallization process for P-2 and P-3 forms resulting in high-purityproduct with the API content preferably above 98% w/w.

In another aspect, the present invention provides other novel crystalforms, exhibiting PXRD patterns denoted as P-4, P-6, P-7 and P-8(alternatively MDT-637 P4, MDT-637 P6, MDT-637 P7 and MDT-637 P8).

In another aspect, the present invention provides a novel amorphousform.

In another aspect, the present invention provides a novel solidcomposite form of API dispersed in a suitable pharmaceutical excipient.

In addition, some novel crystal forms of the present invention areparticularly desirable due to their improved solubility and/orsolid-state stability, producing low dosing levels, ease of handling andprocessing, such as micronization, formulation mixing and blisterfilling, allowing enhanced dosing regimen and administration, lowerdosage as well as the significant reduction and absence of side effectsand toxicity. The novel crystal forms described herein displayunexpected results in terms of their physicochemical and therapeuticproperties.

The present invention comprises a pharmaceutical composition comprisinga therapeutically effective amount of a crystal form of P-3 dihydrate,or P-3 ethanolate, or P-3 monohydrate, or P-3 anhydrous, or P-2anhydrous, P-4, P-6, P-7, P-8, or an amorphous form, or combinationsthereof, and at least one pharmaceutically acceptable carrier.

In addition, the crystal forms and synthesis methods described hereinare improvements over prior art compositions and methods in that theycomprise novel polymorphic forms enabling the production of improvedpharmaceutical formulations and dosage forms having enhanced therapeuticvalue.

Accordingly, it is an object of the present invention to providedisclosure of novel MDT-637 crystal forms and methods of theirpreparation resulting in pharmaceutical products associated withtreatment and prevention of different viral infections, in particularwith treatment and prevention viral infections using respiratory drugdelivery, wherein such forms and compositions are optimized for ease ofdelivery, for dosing, for stability and reduced toxicity.

Accordingly, it is an object of the present invention to provide novelcompositions and methods for alleviating and preventing symptomsassociated with Paramyxovirinae and Pneumovirinae infection.

Another object of the present invention is to provide novel compositionsand methods for alleviating and preventing symptoms associated withmeasles virus, mumps virus, the parainfluenza viruses, and respiratorysyncytial virus (RSV).

Another object of the present invention is to provide novel compositionsand methods for alleviating and preventing symptoms associated withrespiratory syncytial virus (RSV) wherein such symptoms compriserhinitis, otitis media, pneumonia, bronchiolitis and death.

Yet another object of the present invention is to provide novelcompositions and methods for alleviating and preventing symptomsassociated with Paramyxovirinae and Pneumovirinae infection wherein suchcompositions may be delivered in low dosages.

A further object of the present invention is to provide novelcompositions and methods for alleviating and preventing symptomsassociated with Paramyxovirinae and Pneumovirinae infection wherein suchcompositions are optimized for ease of delivery, for dosing, forstability and reduced toxicity.

An additional object of the present invention is to provide novelcompositions and methods for alleviating and preventing symptomsassociated with Paramyxovirinae and Pneumovirinae infection wherein suchcompositions are optimized for ease of delivery to subjects havinglimited ability such as infants, elderly, immunocompromised individualsand subjects having restricted inspiratory airflow.

Another object of the present invention is to provide novel compositionsand methods for alleviating and preventing symptoms associated withParamyxovirinae and Pneumovirinae infection wherein such methodsfacilitate and encourage therapeutic compliance.

Yet another object of the present invention is to provide novelcompositions and methods for alleviating and preventing symptomsassociated with RSV, wherein such compositions are optimized for ease ofdelivery to subjects having restricted inspiratory airflow.

Another object of the present invention is to provide novel compositionsand methods for alleviating and preventing symptoms associated withrespiratory syncytial virus (RSV) wherein such symptoms compriserhinitis, otitis media, pneumonia, bronchiolitis and death and whereinsuch compositions are optimized for ease of delivery, for dosing, forstability and reduced toxicity and wherein such compositions comprisecompositions and methods comprising unique.

A further object of the present invention is to provide novelcompositions and methods comprising unique polymorphic forms of MDT-637and related compounds.

Another object of the present invention is to provide novel compositionsand methods comprising unique polymorphic forms of MDT-637 and relatedcompounds, wherein such compositions comprise the polymorph exhibitingPXRD patterns denoted as P-2, P-3, P-4, P-6, P-7 and P-8.

Another object of the present invention is to provide novel compositionsand methods comprising unique polymorphic forms of MDT-637 and relatedcompounds, and pharmaceutically acceptable excipients.

A further object of the present invention is to provide novelcompositions and methods comprising unique polymorphic forms of MDT-637wherein such polymorphic forms are optimized for delivery to a subjectvia inhalation.

These and other objects, features and advantages of the presentinvention will become apparent after a review of the following detaileddescription of the disclosed embodiments and the appended claims.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 provides a characteristic PXRD pattern of P-2 hydrate crystalform.

FIG. 2 provides a characteristic PXRD pattern of P-2 anhydrous crystalform.

FIG. 3 provides a characteristic FTIR pattern of P-2 hydrate crystalform.

FIG. 4 provides a characteristic DSC trace of P-2 hydrate crystal form.

FIG. 5 provides a characteristic TGA trace of P-2 hydrate crystal form.

FIG. 6 provides a characteristic DVS absorption/desorption curves forconversion between anhydrous and hydrate crystal forms of pattern P-2.

FIG. 7 provides a flow chart showing an example of crystallization,filtration, drying and hydration processes to produce high-purity P-2hydrate crystal form.

FIG. 8 provides a characteristic PXRD pattern of P-3 dihydrate crystalform.

FIG. 9 provides a characteristic PXRD pattern of P-3 monohydrate crystalform.

FIG. 10 provides a characteristic PXRD pattern of P-3 anhydrous crystalform.

FIG. 11 provides a characteristic PXRD pattern of P-3 ethanolate crystalform.

FIG. 12 provides a characteristic FTIR pattern of P-3 dihydrate crystalform.

FIG. 13 provides a characteristic DSC trace of P-3 dihydrate crystalform.

FIG. 14 provides a characteristic TGA trace of P-3 dihydrate crystalform.

FIG. 15 provides characteristic DVS absorption/desorption curves forconversion between anhydrous, monohydrate and dihydrate crystal forms ofpattern P-3 at temperature 20° C.

FIG. 16 provides a flow chart showing an example of re-crystallization,filtration, drying and hydration processes to produce high-purity P-3dihydrate crystal form.

FIG. 17 illustrates the thermodynamic relationship (hierarchy) of theP-2 and P-3 families of crystal forms.

FIGS. 18 a and 18 b provide graphs depicting solubility of differentcrystal forms in simulated lung fluid (SLF): P-2, P-3, and mixture ofP-3 (approximately 75% w/w) and P-2 (approximately 25% w/w). The dataare shown for: (a) non-hydrogenated DPPC (Sigma-Aldrich, USA) and (b)hydrogenated DPPC (Lipoid LLC, USA) surfactants at 0.02% w/vconcentration. Each data point represents a triplicate sample obtainedat temperature 37° C.

FIG. 19 provides a characteristic PXRD pattern of P-4 crystal form.

FIG. 20 provides a characteristic PXRD pattern of P-6 crystal form.

FIG. 21 provides a characteristic PXRD pattern of P-7 crystal form.

FIG. 22 provides a characteristic PXRD pattern of P-8 crystal form.

FIG. 23 gives the results of solubility study of drug concentration instirred suspensions at 25° C. as a function of time for P-2, P-3 formsand their physical mixture in acetonitrile-water 50/50 v/v solution.

FIG. 24 provides a schematic of an inhaler suitable for use with thepresent invention and described for example in Example 27.

FIG. 25 provides a graph showing serial spirometry for high dose levelof MDT-637 P-3 polymorph vs. placebo showing no effect on pulmonaryfunction from inhalation.

FIG. 26 provides a graph showing limited plasma exposure over 10 daysdosing with minimal accumulation between Day 2 and Day 10 across each of3 dose levels of MDT-637 P-3 polymorph.

FIG. 27 provides TABLE 17 showing the inclusion and exclusion criteriaused in selecting patients for Study 1.

FIG. 28 provides TABLE 18 showing the inclusion and exclusion criteriaused in selecting patients for Study 1.

FIG. 29 provides TABLE 19 showing the inclusion and exclusion criteriaused in selecting patients for Study 1.

DETAILED DESCRIPTION OF THE INVENTION

The present invention may be understood more readily by reference to thefollowing detailed description of the specific embodiments includedherein. Reference is made to the accompanying drawings, which form apart hereof, and in which is shown, by way of illustration, variousembodiments of the present disclosure. Although the present inventionhas been described with reference to specific details of certainembodiments thereof, it is not intended that such details should beregarded as limitations upon the scope of the invention.

The entire text of the references mentioned herein are herebyincorporated in their entireties by reference including U.S. ProvisionalApplication Ser. No. 61/666,258 filed Jun. 29, 2012, U.S. Pat. No.6,495,580 filed on Jan. 29, 1999, U.S. patent application Ser. No.10/524,162 filed on Aug. 11, 2003 and U.S. patent application Ser. No.10/524,313 filed on Aug. 11, 2003.

The present invention provides novel polymorphs of the compound MDT-637described in U.S. Pat. No. 6,495,580 and its structure is shown below.The polymorphs described herein are novel and unobvious in the way thattheir unique crystalline forms do not follow or result from any priorart and/or theoretical computations/predictions. The polymorphsdescribed herein are produced under specific unobvious crystallizationconditions which do not follow from any prior art chemical synthesisproduction steps or procedures: crystallization from the final syntheticstep may result in different forms or mixture of different forms withoutappropriate controls disclosed herein. An additional point of novelty isthat crystal forms discovered have unexpected and unique physiochemicalproperties such as solubility, dissolution rate, stability, chemicalreactivity, hygroscopicity, and powder handling properties which provideadvantageous characteristics enabling the production of improvedpharmaceutical formulations and dosage forms having enhanced therapeuticvalue.

Though not wishing to be bound by the following theory, it is believedthat MDT-637, its isomers and polymorphs function as fusion inhibitors.For example, in the case of RSV, it is believed that MDT-637, itsisomers and polymorphs function as a fusion inhibitor that prevents theattachment of RSV to human cells via F fusion and G attachmentglycoproteins.

As used herein, the phrases and terms “active pharmaceuticalingredient”, “API”, “drug substance” or “drug” refer to MDT-637 compoundwith the chemical name: phenol,2,2′-[(4-hydroxyphenyl)methylene]bis[4-[[(5-methyl-1H-tetrazol-1-yl)imino]methyl]];alternate name:5,5′-Bis[1-(((5-methyl-1-H-tetrazolyl)imino)methyl)]-2,2′,4″-methylidynetrisphenol; molecular formula C₂₅H₂₂N₁₀O₃) as described by the followingstructure:

As used herein, the terms “crystal form of pattern P-2, or P-3, or P-4,or P-6, or P-7, or P-8” or “form P-2, or P-3, or P-4, or P-6, or P-7, orP-8” or MDT-637 P2, MDT-637 P3, MDT-637 P4, MDT-637 P6, MDT-637 P7,MDT-637 P8, all refer to the designation of different polymorphs andsolvates of the API according to their PXRD patterns, sequentiallynumbered according to the time of their discovery. Several crystalforms, such as isomorphic solvates, may exhibit a similar PXRD pattern.

As used herein, the terms “Characteristic PXRD pattern” or“characteristic FTIR pattern” means that these patterns exhibit the samepositions and sequence of peaks within the limits defined by theanalytical methods and instrumentation. Because of the variation betweendifferent samples, instruments and natural variability of measurements,the peak positions may deviate from reported positions. In case of thePXRD, this deviation may be as much as 0.2 degrees in 2θ values. Theremay also be large differences in PXRD peak intensities due to instrumentand sample variability, particle size, crystallinity, and the phenomenonof preferential crystal orientation known in the prior art.

As used herein, the terms “characteristic DSC trace” or “characteristicTGA trace” or “characteristic DVS curve” refer to the shape ofcorresponding dependencies such as the magnitude of variation of theparameter measured as a function of temperature and relative humidity (%RH), as well as to the major inflection points in such dependencies. Itis understood in the prior art that these dependences may exhibitsignificant deviations due to the variation between different samplesand measurement technique, in particular, variation in the level ofinitial sample hydration, the type and scanning speed of the instrumentsutilized.

As used herein, the term “crystallinity” generally describesimperfections of crystal lattice (a multitude of crystal defects) thatcan generally be associated with broadening of the PXRD peaks.

As used herein, the term “solid composition” generally refers to the APIdistributed in a solid matrix of a suitable pharmaceutical excipient,both in a form of molecular dispersion, amorphous dispersion ornanoparticles.

As used herein, the term “water-miscible solvent” generally comprises asolvent that can be mixed in any ratio with drug solution without phaseseparation.

As used herein, the term “water-immiscible solvent” generally comprisesa solvent that can be mixed only partially with drug solution withoutphase separation.

As used herein, the term “antisolvent” generally comprises a solventthat can be used in crystallization of the drug compound which ismiscible with drug solution but in which the drug is practicallyinsoluble.

As used herein, the term “ambient conditions” generally refers to arange of temperatures typically between 15-37° C., and relative humiditybetween 40-100%.

As used herein, the terms “pharmaceutically acceptable excipients orpharmaceutically acceptable carrier medium” comprise any solid or liquidsubstances, diluents, dispersion or suspension agents, surface activeagents, isotonic agents, thickening or emulsifying agents,preservatives, solid binders, lubricants and the like, as suited to theparticular dosage form desired. Remington's Pharmaceutical Sciences,Fifteenth Edition, E. W. Martin (Mack Publishing Co., Easton, Pa., 1975)discloses various carriers used in formulating pharmaceuticalcompositions and known techniques for the preparation thereof. Exceptinsofar as any conventional carrier medium is incompatible with the APIof the invention, such as by producing any undesirable biological effector otherwise interacting in a deleterious manner with any othercomponent(s) of the pharmaceutical composition, its use is contemplatedto be within the scope of this invention.

Crystal (polymorphic) forms of the same drug substance can be defined as“different crystalline forms of the same pure substance in which themolecules have different arrangements and/or different conformations ofthe molecules” (Grant, D. J. W., Theory and Origin of Polymorphism, inPolymorphism in Pharmaceutical Solids, H. G. Brittain, Editor 1999,Marcel Dekker: New York). However the regulatory agencies which controlor monitor drug products in different countries, and pharmaceuticalindustry in general, define solid polymorphism broadly. This definitionincludes all crystalline forms that contain a drug substance, includingsolvates with different stoichiometric and non-stoichiometricrelationships, as well as salts and amorphous materials (U.S. Departmentof Health and Human Services, F.D.A., Center for Drug Evaluation andResearch (CDER), ANDAs: Pharmaceutical Solid Polymorphism, Chemistry,Manufacturing, and Controls Information, in Guidance for Industry 2007).As used herein, “forms” or “polymorphs” or “crystal polymorphic forms”are understood to be different crystalline forms in which the moleculeshave different arrangements and/or different conformations of themolecules, including forms of a pure substance and all crystalline formsthat contain a drug substance, including solvates with differentstoichiometric and non-stoichiometric relationships, as well as saltsand amorphous materials.

The majority of molecules of synthetic and semi-synthetic origin, whichare currently in pharmaceutical development, have high molecular weightand significant conformational mobility. Such molecules may havemultiple polymorphic forms and numerous solvates. Crystal polymorphismhas a direct effect on several characteristics of both drug substances(active pharmaceutical ingredients, APIs) and solid drug products. Forexample, polymorphism may affect API physicochemical properties such asmelting point, intrinsic density, hardness, hygroscopicity; powdercharacteristics such as bulk density, flowability, cohesiveness and mayalso differ in analytical characteristics such as powder X-raydiffraction (PXRD) pattern and/or spectroscopic pattern, measured forexample using Fourier-Transform Infra-Red (FTIR) spectroscopy or byNuclear Magnetic Resonance (NMR) spectroscopy, and thermal behavior asmeasured using differential scanning calorimetry (DSC) andthermogravimetric analysis (TGA), as well as water sorption/desorptionprofile as measured using Dynamic Vapor Sorption (DVS) method.Importantly, polymorphism affects the equilibrium solubility anddissolution rates in different solvents (including bodily fluids), andtherefore may produce different pharmacokinetic profiles and therapeuticconcentrations for the same drug molecule. Unstable or metastablepolymorphs may convert into more thermodynamically stable polymorphsduring storage or processing. Polymorphs of the same drug molecule mayalso exhibit different chemical impurities and degradation productsafter processing and upon storage. Thus different polymorphs may haveadvantageous and disadvantageous properties for drug products. Thestructure, properties and methods of preparations of new polymorphs ofMDT-637 are subjects of the present invention.

Powder X-ray diffraction (PXRD) analysis was used as the major techniquefor crystal form identification and performed by the methods known inthe art using Bruker D8 Advance Diffractometer with Bragg-Brentanooptics. A conventional CuK_(α), radiation of wavelength 1.5418 {acuteover (Å)} was generated with settings of 40 kV and 40 mA throughout allanalyses. Samples were mounted onto a silicon low-background holder andflattened with wax paper and a glass slide to form a smooth, thin,uniform sample surface. The data was collected from 3-40° 2θ values insteps of 0.01° 2θ and at a rate of 0.3 seconds/step. Diffractionpatterns were processed with Eva 13 software (Bruker AXS GmbH,Karlsruhe, Germany).

As described below, several novel polymorphic forms and solvates of theAPI were discovered. The materials of patterns P-2 and P-3 showparticular utility with regard to stability, solubility and suitabilityfor development of novel pharmaceutical dosage forms. These specificpolymorphs are highly relevant to the final API production step and havea high impact on the product manufacturability, e.g., purity,micronization, powder blending, dose filling and formulationperformance, including but not limited powder potency, contentuniformity and aerosolization efficiency (delivered dose and aerodynamicparticle size distribution as measured by a Cascade Impactor (CI), suchas Andersen Cascade Impactor (ACI) and New Generation Impactor (NGI)techniques known to those skilled in the art.

The characteristic diffraction peaks of the hydrate crystal form ofpattern P-2 are shown in FIG. 1 and listed in TABLE 1. The mostcharacteristic peaks for P-2 hydrate are observed at 2θ values: 4.83°;8.42°; 9.61°; 11.83°; 14.60°; 16.94°; 19.25°; 20.46°; 24.09°; 24.85°;27.76°; 28.56°.

TABLE 1 P-2 (Hydrate Crystal Form) Characteristic Diffraction PeaksRelative 2Θ, ° d-value, Å Intensity % 4.83 18.30 12.50 6.83 12.92 2.008.42 10.50 22.60 9.61 9.19 78.70 11.83 7.47 43.30 13.84 6.39 5.00 14.606.06 100.00 15.86 5.58 5.50 16.35 5.42 8.40 16.94 5.23 16.80 17.76 4.992.40 18.53 4.78 6.60 19.25 4.61 14.30 20.46 4.34 15.50 21.31 4.17 7.1022.13 4.01 4.00 22.98 3.87 6.80 24.09 3.69 33.00 24.85 3.58 10.60 25.503.49 5.10 25.80 3.45 7.80 27.00 3.30 4.30 27.76 3.21 15.60 28.56 3.1217.00 29.71 3.00 5.20 30.24 2.95 4.80 31.25 2.86 0.60 32.24 2.77 4.3033.99 2.64 2.50 35.36 2.54 1.70 36.01 2.49 1.90

The characteristic diffraction peaks of the anhydrous crystal form ofpattern P-2 are shown in FIG. 2 and listed in TABLE 2. The mostcharacteristic peaks for P-2 anhydrous form are observed at 2θ values:8.58°; 10.64°; 12.58°; 14.51°; 15.89°; 17.20°; 21.06°; 21.41°; 22.37°;25.43°; 27.62°.

TABLE 2 P-2 (Anhydrous Crystal Form) Characteristic Diffraction PeaksRelative 2Θ, ° d-value, Å Intensity % 5.35 16.52 2.20 6.81 12.97 2.708.58 10.30 9.50 10.64 8.31 100.00 12.58 7.03 11.90 13.52 6.54 3.00 14.516.10 79.30 15.89 5.57 20.30 17.20 5.15 55.50 18.85 4.70 6.20 20.18 4.404.70 21.06 4.22 16.50 21.41 4.15 19.20 22.37 3.97 25.50 23.26 3.82 5.6024.68 3.60 10.00 25.43 3.50 31.00 26.22 3.40 10.60 27.62 3.23 15.4028.92 3.08 3.80 29.82 2.99 8.10 31.95 2.80 7.50 33.30 2.69 2.10

The PXRD data for P-2 forms obtained under controlled temperature andhumidity indicated a pronounced difference between diffraction patternsfor hydrated and anhydrous forms of P-2 attributed to lattice expansionand water incorporation. Therefore it is possible to distinguish thehydrated and anhydrous forms of P-2 lattice from the PXRD peak shiftsalthough the lattice does not undergo dramatic structural changes.

FIG. 3 represents a characteristic FTIR pattern of the P-2 hydrate. Themost characteristic absorption bands in the region are between 800-1200cm⁻¹, corresponding to P-2 crystal forms are: 836.5 cm⁻¹; 980.9 cm⁻¹,double peak at 1080.0 and 1091.9 cm⁻¹, 1208.9 cm⁻¹, 1238.6 cm⁻¹ and1287.67 cm⁻¹.

DSC trace for P-2 hydrate (FIG. 4) indicate a sharp melting peak atapproximately 263° C., with a small thermal event between 40° C. to 60°C., likely dehydration as confirmed by loss of weight within thistemperature interval according to TGA data (FIG. 5). Though not wishingto be bound by any particular theory, it is believed that P-2 loseswater relatively easily at temperatures about 60° C., but melts withoutlattice transition and may constitute a channel hydrate with a stabledehydrate product (stable crystal lattice without further phasetransition).

Assessment of the critical water activity using small equilibrationsteps (1% RH) with DVS measurements of sorption and desorption isotherms(FIG. 6) shows that this activity for transition between P-2 hydrate andanhydrous forms is typically between 30-40% RH. The characteristicgravimetric loss of water is between 1-4.5% w/w.

The manufacture of specific forms herein was controlled through theunderstanding of their thermodynamic relationship (phase diagram) andthrough the kinetics of crystallization process. The high puritymaterial of pattern P-2 hydrate was recrystallized from P-3 ethanolateform dissolved in acetone/water mixture using water as an antisolventwith parallel cooling crystallization as disclosed in FIG. 7. Theproduct was filtered, washed and dried under controlled conditions. Theabsence of ethanol was determined by ¹H NMR analysis. The chemicalpurity of the material produced was determined using an HPLC method andpreferable above 99.0%.

P-2 anhydrous form can be obtained from the P-2 hydrate by dehydratingthis material below 30% RH and/or by heating it above 60° C.

The characteristic diffraction peaks of the dihydrate crystal form ofpattern P-3 are shown in FIG. 8 and listed in TABLE 3. The mostcharacteristic peaks for P-3 dihydrate are observed at 2θ values: 7.03°;8.16°; double peak at 9.47° and 9.75°; 11.60°; 14.24°; 17.95°, 19.64°;20.26°; 24.52°; 26.24°; 27.25°. All crystal forms of pattern P-3 areisomorphic and therefore their PXRD patterns are similar and thecorresponding peak values are very close to each other for P-3 dihydrate(FIG. 8, TABLE 3), monohydrate (FIG. 9; TABLE 4); anhydrous (FIG. 10;TABLE 5) as well as ethanolate (FIG. 11; TABLE 6) crystal forms. Asdiscussed before, the peak position is affected by the accuracy of thePXRD instrument, sample preparation and by the peak fitting procedure.Therefore a deviation in measured peak positions is expected. However aspecific PXRD pattern can easily be recognizable by one skilled in theart by considering peak positions together with peak intensities andcorrelating one PXRD pattern to a reference pattern.

TABLE 3 P-3 (Dihydrate Crystal Form) Characteristic Diffraction PeaksRelative 2Θ, ° d-value, Å Intensity % 4.77 18.52 3.20 7.03 12.57 4.808.16 10.83 100.00 9.47 9.33 24.20 9.75 9.07 10.80 11.60 7.63 30.90 13.386.61 5.20 13.64 6.49 4.90 14.24 6.22 40.30 15.96 5.55 8.60 16.37 5.417.20 16.94 5.23 1.30 17.95 4.94 24.70 18.47 4.80 7.20 19.08 4.65 3.0019.64 4.52 19.50 20.26 4.38 13.50 20.96 4.24 3.60 21.48 4.13 3.40 22.124.01 5.40 22.64 3.92 2.40 23.19 3.83 9.30 23.93 3.71 3.50 24.17 3.686.20 24.52 3.63 15.20 25.26 3.52 9.50 26.24 3.39 15.40 27.25 3.27 6.8027.54 3.24 6.40 28.52 3.13 2.80 29.12 3.06 1.90 29.46 3.03 2.40 29.962.98 1.80 31.22 2.86 1.50

TABLE 4 P-3 (Monohydrate Crystal Form) Characteristic Diffraction PeakRelative 2Θ, ° d-value, Å Intensity % 4.80 18.38 3.10 7.06 12.51 3.108.25 10.71 87.10 9.55 9.26 64.50 9.84 8.98 12.80 11.66 7.58 53.50 13.586.51 5.10 14.30 6.19 100.00 16.02 5.53 8.60 16.47 5.38 8.60 17.05 5.203.20 18.03 4.92 41.50 18.54 4.78 7.30 19.13 4.64 8.90 19.70 4.50 35.6020.29 4.37 37.80 21.00 4.23 5.30 21.54 4.12 7.70 22.24 3.99 5.90 22.663.92 3.80 23.25 3.82 7.80 24.02 3.70 8.20 24.55 3.62 16.90 25.29 3.5216.10 26.31 3.39 14.80 27.26 3.27 30.30 27.63 3.23 7.40 28.56 3.12 7.3029.17 3.06 3.30 29.53 3.02 4.40 31.25 2.86 3.50

TABLE 5 P-3 (Anhydrous Crystal Form) Characteristic Diffraction PeaksRelative 2Θ, ° d-value, Å Intensity % 4.77 18.52 3.20 7.08 12.48 2.808.27 10.68 88.70 9.54 9.27 71.40 9.86 8.96 13.40 11.68 7.57 58.30 13.736.44 8.00 14.29 6.19 100.00 15.96 5.55 7.30 16.38 5.41 5.80 16.62 5.337.30 18.07 4.90 34.70 18.51 4.79 6.90 19.09 4.65 8.00 19.79 4.48 32.3020.30 4.37 34.20 20.97 4.23 5.20 21.58 4.11 7.80 22.23 4.00 6.30 22.633.93 5.20 23.25 3.82 8.00 23.98 3.71 8.50 24.54 3.62 15.90 25.27 3.5215.30 26.36 3.38 15.60 27.25 3.27 28.30 27.67 3.22 7.50 28.51 3.13 5.6029.13 3.06 3.50 29.57 3.02 3.70 30.10 2.97 3.30 31.20 2.86 3.50

TABLE 6 P-3 (Ethanolate Form) Characteristic Diffraction Peaks Relative2Θ, ° d-value, Å Intensity % 7.16 12.34 5.80 8.25 10.71 100.00 9.54 9.2627.40 9.85 8.98 10.80 11.69 7.57 27.80 13.47 6.57 6.20 13.74 6.44 6.2014.32 6.18 44.80 15.99 5.54 11.30 16.45 5.39 10.50 18.06 4.91 22.2018.51 4.79 9.40 19.16 4.63 4.30 19.73 4.50 15.60 20.33 4.36 11.10 20.974.23 3.10 21.56 4.12 4.60 22.17 4.01 8.00 22.67 3.92 3.50 23.27 3.828.20 24.19 3.68 6.20 24.54 3.62 10.70 25.29 3.52 8.60 26.27 3.39 15.5027.32 3.26 6.70 27.67 3.22 5.60 28.57 3.12 2.00 29.19 3.06 1.90 29.463.03 2.10 30.06 2.97 1.20 31.26 2.86 1.50

FIG. 12 represents a characteristic FTIR pattern of the P-3 hydrate. Themost characteristic absorption bands in the region are between 800-1200cm⁻¹, corresponding to P-3 crystal forms are: 831.5 cm⁻¹; 994.9 cm⁻¹,1092.0 cm⁻¹, 1211.0 cm⁻¹, 1237.7 cm⁻¹ and 1290.5 cm⁻¹.

DSC trace for P-3 dihydrate (FIG. 13) exhibits a wide endothermic eventbetween 220° C. to 280° C., the characteristic delta-shaped peak ofwhich suggests several thermal events, such asrecrystallization/transition between different forms, in this interval.TGA data shows weight loss of 5-7% between 30° C. to 100° C. for thismaterial (FIG. 14). Therefore P-3 dihydrate loses water at a highertemperature than P-2 hydrate but undergoes some additional changes afterthis event and the melting peak may correspond to a different form.

Assessment of the critical water activity using small equilibrationsteps (1% RH) with DVS measurements of sorption and desorption isothermsfor P-3 forms (FIG. 15) shows two transition points (mono- anddihydrate) at approximately 3 and 13% RH. The characteristic gravimetricloss of water for P-3 dihydrate is between 5.5-8.5% w/w. The profounddifference of the DVS curves between the P-2 and P-3 forms can beutilized as a secondary technique for quantitative phase analysis ofthese forms.

P-3 ethanolate was obtained by crystallization from solution containingsufficient amount of ethanol above a certain critical concentration.This critical concentration in acetone/water/ethanol system, used bothfor the API purification and generation of P-3 ethanolate form, wasfound to be approximately 33% v/v. A reproducible crystallizationprocess was achieved at ethanol concentrations above this limit. FIG. 16illustrate the process for production of high purity P-3 dihydrate whichis obtained by recrystallization of P-3 ethanolate dissolved inacetone/water mixture using ethanol as an antisolvent with parallelcooling crystallization, following by filtration, washing, hydration anddrying steps under controlled conditions. The absence of ethanol isdetermined by ¹H NMR analysis. The chemical purity of the materialproduced is determined using an HPLC method and preferably above 99.0%.

P-3 monohydrate and P-3 anhydrous forms can be obtained from the P-3dihydrate by drying this material below approximately 13% RH formonohydrate and below approximately 3% RH for anhydrous form and/or byprolonged heating it at elevated temperatures preferably above 60° C.

It was discovered that P-2 dihydrate is the most stable crystal formunder ambient conditions (i.e. temperatures preferably between 15-37°C., relative humidity preferably between 40-100%). It was alsodiscovered that P-3 ethanolate is the most stable crystal form withsufficient activity of ethanol (sufficiently high concentration ofethanol in solution) preferably above 0.3. FIG. 17 shows a diagram ofthermodynamic relationship between different P-2 and P-3 forms.

Time-dependent solubility study was carried out in simulated lung fluid(SLF) containing the surfactant dipalmitoylphosphatidylcholine (DPPC).The DPPC is a major surfactant present in the human lung. Since MDT-637is formulated for delivery through nasal passage to the lung, it is veryimportant to have the API solubility assessed in SLF modified with 0.02%DPPC as such information is useful in understanding the drugfunctionality in the lung. FIG. 18 shows results of dissolution studiescarried out for 5 days with two different DPPC surfactants atconcentration 0.02% w/v, and with P-2 hydrate, P-3 dihydrate and mixedP-3/P-2 material at 37° C. The data indicates that the solubility of P-3dihydrate is approximately 550 ng/mL (1077 nM) versus 100 ng/mL (196 nM)for P-2 hydrate. Solubility in pure water, also assessed during thesestudies for both P-2 and P-3 forms, is significantly lower: it is belowthe HPLC limit of detection, approximately equivalent to 6 nM. Theseresults indicate, firstly, that P-2 form is approximately a factor of 5to 6 lower than solubility of P-3 dihydrate.

In addition, P-3 dihydrate material was stable against solid-statechanges and chemical degradation in open-dish stability studies at 40°C./75% RH for 4 weeks as shown in the Examples.

P-3 dihydrate material was also stable against solid-state conversion informulation with lactose while stored in sealed glass bottles for 39weeks at 40° C./75% RH as shown in the Examples.

Furthermore, micronization did not affect the crystallinity, phasepurity or induce any chemical degradation for P-3 dihydrate as shown inthe Examples.

Following the experiments discussed above, it was therefore discoveredthat P-3 dihydrate, despite being a less thermodynamically stable formthan P-2 hydrate, is kinetically stable to solid-state conversions,dehydration and chemical degradation, and possesses the advantage ofsignificantly higher solubility than P-2 hydrate form. P-3 dihydrate istherefore a sufficiently robust and suitable crystal form forpharmaceutical development. Comparison of some major physicochemicalproperties of P-3 dehydrate versus P-2 hydrate is presented in TABLE 7.

TABLE 7 Characteristics Property Method P-2 hydrate P-3 dihydrateAppearance N/A white solid white solid Crystal habit Microscopy acicularacicular Melting point DSC 263° C. range: 220-280° C. Characteristicrelative DVS 30-40% 3-17% humidity of dehydration (20° C.) Water contentStoichiometric 3.3 wt % (monohydrate) 6.6 wt % Crystal (skeletal)Nitrogen Pycnometry g/cm³ 1.61 g/cm³ density Characteristic SympatecHELOS & 1.1 μm 1.5 μm medium volume RODOS particle size (micronized)Specific surface area BET 22.6 m²/g 11.0 m²/g Poured bulk powderVolumetric 0.16 g/cm³ 0.22 g/cm³ density Tapped bulk powder Volumetric0.19 g/cm³ 0.30 g/cm³ density Hausner ratio Volumetric 1.20 1.38 Carrindex Volumetric 16.7% 27.5%

As recognized by those skilled in the art, the polymorphic form chosenis typically the most thermodynamically stable form because highersolubility or superior powder characteristics of some metastable formsusually do not justify the regulatory risks associated withuncontrollable conversion of such forms in the final drug product,either during their processing and/or storage. However, in contrast towhat would be expected, the inventors herein selected the lessthermodynamically stable form, P-3 dihydrate for a preferred embodiment,considering it to represent a more viable option for product developmentas discussed above and in the Examples. Although development of a lessstable solid form is not advisable, the risk of multifold reduction insolubility, and potentially drug concentration against the clinicalstrains of RSV is significant.

In addition to the crystal forms of the P-2 and P-3 patterns, othernovel crystal forms were discovered, these include but are not limitedto, P-4, P-6, P-7 and P-8.

The characteristic diffraction peaks of crystal form of pattern P-4 areshown in FIG. 19 and TABLE 8. The most characteristic peaks for P-4 formare observed at 2θ values: 4.31°, 7.99°, 9.37°, 11.02°, 13.04°, 13.43°,14.18°, 16.13°, 16.70°, 17.08°, 17.42°, 17.92°. This form was obtainedfrom saturated DMF solution using cooling, evaporation or antisolventcrystallization procedures. Though not wishing to be bound by anyparticular theory, it is believed that P-4 crystal form is likely a DMFsolvate.

TABLE 8 P-4 (Crystal Form) Characteristic Diffraction Peaks PatternRelative 2Θ, ° d-value, Å Intensity % 4.31 20.49 22.00 7.03 12.57 5.407.99 11.06 100.00 8.92 9.90 3.00 9.37 9.43 16.60 11.02 8.02 8.90 13.046.78 9.40 13.43 6.59 11.60 13.69 6.46 6.70 14.18 6.24 12.60 14.76 6.003.80 16.13 5.49 15.10 16.70 5.30 21.00 17.08 5.19 17.50 17.42 5.09 51.6017.92 4.95 8.90 18.84 4.71 7.00 19.18 4.62 7.00 20.50 4.33 11.30 22.393.97 1.50 22.99 3.87 2.50 24.09 3.69 2.70 25.07 3.55 4.70 25.61 3.485.20 27.40 3.25 5.00 28.30 3.15 10.40 29.49 3.03 1.40

The characteristic diffraction peaks of crystal form of pattern P-6 areshown in FIG. 20 and TABLE 9. The most characteristic peaks for P-6 formare observed at 2θ values: double peak at 3.43° and 3.89°, 6.89°, doublepeak at 7.87° and 8.25°, 10.88°, 13.06° and 13.81°, 16.12° 17.38° and18.51°. This form was obtained from saturated DMF solution usingantisolvent crystallization with MTBE.

TABLE 9 P-6 (Crystal Form) Characteristic Diffraction Peaks PatternRelative 2Θ, ° d-value, Å Intensity % 3.43 25.71 100.00 3.89 22.69 53.206.89 12.83 4.70 7.87 11.23 18.10 8.25 10.72 8.20 9.31 9.49 1.90 9.928.91 1.60 10.88 8.13 3.20 13.06 6.78 2.80 13.81 6.41 4.00 16.12 5.4911.10 17.38 5.10 37.50 18.51 4.79 9.20 20.25 4.38 2.30 22.02 4.03 1.5022.83 3.89 2.20 25.23 3.53 5.90 27.52 3.24 4.80 28.18 3.16 3.60

The characteristic diffraction peaks of crystal form of pattern P-7 areshown in FIG. 21 and TABLE 10. The most characteristic peaks for P-7form are observed at 2θ values: 5.18°, 6.59°, 7.70°, double peak 10.02and 10.62°, double peak at 12.18° and 12.50°, 15.16°, double peak at15.88° and 16.4°, double peak at 17.20 and 17.42°, double peak at 18.07°and 18.25°. This form was obtained from saturated NMP solution byantisolvent precipitation in two-phase ternary systemNMP-cyclohexane-water.

TABLE 10 P-7 (Crystal Form) Characteristic Diffraction Peaks PatternRelative 2Θ, ° d-value, Å Intensity % 5.18 17.03 37.60 6.59 13.40 100.007.70 11.47 7.50 10.02 8.82 46.00 10.62 8.33 30.20 12.18 7.26 28.00 12.507.07 17.40 13.40 6.60 6.50 15.16 5.84 59.50 15.88 5.58 73.40 16.40 5.4013.00 17.20 5.16 32.00 17.42 5.09 90.70 18.07 4.91 27.30 18.25 4.8620.00 19.57 4.53 4.80 20.26 4.38 13.70 21.35 4.16 15.40 21.94 4.05 30.7022.76 3.90 10.00 23.34 3.81 24.80 24.02 3.70 8.90 24.46 3.64 9.90 25.253.52 18.00 26.11 3.41 9.30 26.70 3.34 6.90 28.11 3.17 13.30 28.87 3.094.40 29.35 3.04 1.90 29.86 2.99 1.80 30.69 2.91 3.20 31.90 2.80 2.2033.07 2.71 1.00 35.46 2.53 2.20

The characteristic diffraction peaks of crystal form of pattern P-8 areshown in FIG. 22 and TABLE 11. The most characteristic peaks for P-8form are observed at 2θ values: 4.15°, 7.85°, 9.33°, 14.29°, triple peakat 15.84, 16.68 and 17.14°, 18.53°, 20.27°, 23.90°, 24.80° and 27.39°.This form was obtained from two different crystallization systems:first, antisolvent precipitation from saturated DMSO solution withaddition of water and, second, by antisolvent precipitation fromsaturated DMF solution using acetone/water mixture as the antisolvent.

TABLE 11 P-8 (Crystal Form) Characteristic Diffraction Peaks PatternRelative 2Θ, ° d-value, Å Intensity % 4.15 21.30 18.40 6.89 12.81 3.307.85 11.25 65.30 9.33 9.47 30.90 11.67 7.58 15.20 12.86 6.88 4.30 14.296.20 100.00 15.84 5.59 22.80 16.68 5.31 50.00 17.14 5.17 69.10 18.534.78 12.10 20.27 4.38 21.50 21.06 4.21 8.90 22.00 4.04 6.50 22.72 3.918.90 23.90 3.72 29.80 24.80 3.59 16.10 27.39 3.25 21.50 28.32 3.15 9.8030.03 2.97 3.20 31.83 2.81 3.50 35.22 2.55 2.70

The PXRD pattern of the amorphous form does not exhibit characteristicdiffraction peaks but show “halos” with typically one or more maxima.The position of these maxima may vary depending on the preparationtechnique used for the amorphous material. The amorphous form can beobtained by several techniques including but not limited to: (a) rapidcooling of saturated API solution; (b) fast evaporation of APIsolutions; (c) fast antisolvent precipitation; (d) spray-drying and (b)freeze-drying. Examples are provided below.

“Solid composition” is yet another embodiment of the API. Similar to theamorphous form, solid composition may not produce a characteristic PXRDpattern identifiable with specific crystalline API diffraction peaks.One important aspect of this invention is the preparation methodologyfor the API: in certain embodiments, an excipient matrix, in the form ofsolid composite microparticles is used wherein the drug is blended intouniform solid phase with selected excipients. The excipients areselected in order to optimize various parameters, including but notlimited to, drug dissolution rate, enabling mucoadhesive propertiesafter the delivery of microparticles into biospaces, combining antiviralproperties of excipients synergistically (to provide a more significantand/or prolonged therapeutic antiviral effect greater than that achievedby the API alone).

The processes for production of solid compositions include but notlimited to several techniques: (a) rapid co-precipitation cooling duringmixing of saturated API solution with saturated excipient solution; (b)fast evaporation of suitable API-excipient solutions; (c) spray-dryingof suitable API-excipient solutions, (d) freeze-drying of suitableAPI-excipient solutions. Examples are provided below.

Solid compositions or formulations of the API with selected excipientswhich form composite solid microparticles, or films, or compositematrixes, where the drug and excipient are physically or chemicallyattached to each other within composite phase, whereas the excipientscan be small molecules or macromolecular substances, including but notlimited to different forms and chemical modifications of lactose,trehalose, sugars, mannitol, amino-acids, polymers including but notlimited to different chemical modifications of hydroxypropylmethylcellulose (HPMC) and microcrystalline cellulose with sodiumcarboxymethylcellulose (e.g. Avicel CL-611, RC-581, RC-591 availablefrom FMC Inc. Philadelphia, Pa.), naturally occurring polysaccharidesextracted from red seaweed (carrageenans) supplied by FMC Inc. asGELCARIN™, VISCARIN™, and SEASPEN PF™ carrageenans and differentdispersing and wetting agents such as polysorbate 80, antioxidants (e.g.ascorbic acid, sodium ascorbate, sodium bisulfate, disodiumethylenediamine tetraacetate (EDTA) and osmolarity modifiers (e.g.dextrose).

In comparison to currently available therapeutics, the MDT-637 compoundsdescribed herein display significantly enhanced efficacy. The keyefficacy features uniquely associated with the compounds include:reduction in hospitalization stay and progression to intensivecare/ventilator, reduction in symptom duration and reduction inrespiratory distress index. In addition, the compounds and compositionsof the present invention are also effective against drug resistantstrains of RSV, i.e. strains resistant to SYNAGIS® (palivizumab).

As described in the examples, and in particular Example 27, experimentsand phase 1 clinical programs evaluating the safety of the MDT-637compounds have been successfully completed. MDT-637 polymorphs such asMDT-637 P3 were determined to be safe and well tolerated in three dosesup to 132 mcg TID for 10 days. In a first randomized double-blindplacebo controlled study (Study 1), where MDT-637 was administered tohealthy volunteers in single ascending doses, no clinically significantchanges in pulmonary function, ECG, laboratory values vitals or physicalexamination were observed.

In a second study (Study 2) also designed to assess safety andtolerability MDT-637 was administered to healthy volunteers for tendays. Ascending doses were tested from 66 mcg twice daily to 132 mcgthree times daily for ten days in approximately forty volunteers. Thestudy findings demonstrated that MDT-637 was safe and well tolerated atdoses up to 132 mcg three times daily for ten days; in addition nosignificant changes were observed in pulmonary function or reports ofpulmonary adverse events.

In a third study (Study 3) further designed to assess safety andtolerability ascending doses of MDT-637 were administered to subjectswith intermittent or mild to moderate asthma. Ascending doses weretested from 66 mcg once daily to 132 mcg three times daily in aapproximately ten volunteers. The study findings demonstrated noclinically significant changes in pulmonary function or reports ofpulmonary adverse events.

The API formulations or compositions with the discovered crystal formsmay be prepared in various forms for administration, including powders,pellets, tablets, caplets, pills or dragées, or can be filled insuitable containers, such as capsules, blisters or, in the case ofsuspensions, filled into bottles or vials. Methods of administration ofsuch formulations are well known to those skilled in the art, andinclude but are not limited to delivery via facemask nebulizers,facemask inhalers, inhalers, nebulizers and variations thereof.

Pharmaceutical organic or inorganic solid or liquid carrier mediasuitable for enteral or parenteral administration can be used to make upthe composition. Gelatine, lactose, starch, magnesium, stearate, talc,vegetable and animal fats and oils, gum, polyalkylene glycol, or otherknown carriers or excipients for medicaments may all be suitable ascarrier media.

Preferably the API crystal forms can be prepared for pulmonary or nasaldrug delivery using different respiratory formulations withpharmaceutically acceptable excipients such as lactose in its amorphousand crystalline forms and/or dispersed in suitable excipient mediamatrix as nanoparticles to enable more efficient drug delivery.

In the pharmaceutical compositions of the invention, the active agentmay be present in any therapeutically effective amount, which istypically at least 0.1% and generally not more than 90% by weight, basedon the total weight of the composition, including carrier medium and/orsupplemental active agent(s), if any. Preferably, the proportion ofactive agent varies between 1-50% by weight of the composition.Depending on the mode of administration, the pharmaceutical compositionwill comprise from 0.05 to 99% by weight, preferably from 0.1 to 70% byweight, more preferably from 0.1 to 50% by weight of the activeingredient, and, from 1 to 99.95% by weight, preferably from 30 to 99.9%by weight, more preferably from 50 to 99.9% by weight of apharmaceutically acceptable carrier, all percentages being based on thetotal weight of the composition.

An appropriate dosage level will generally be about 0.001 to 1000 mg perkg patient body weight per day and can be administered in single ormultiple doses. In various aspects, the dosage level will be about 0.01to about 500 mg/kg per day, about 0.1 to 250 mg/kg per day, or about0.05 to 100 mg/kg per day. A suitable dosage level can be about 0.001 to1000 mg/kg per day, about 0.01 to 500 mg/kg per day, about 0.01 to 250mg/kg per day, about 0.05 to 100 mg/kg per day, or about 0.1 to 50 mg/kgper day. Within this range the dosage can be 0.05 to 0.5, 0.5 to 5.0 or5.0 to 50 mg/kg per day. For inhalant administration, the compositionsare provided in a micronized or inhalant composition containing 0.01 to1000 milligrams of the active ingredient, particularly 0.01, 0.1 1.0,5.0, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 400, 500, 600,750, 800, 900 and 1000 milligrams of the active ingredient for thesymptomatic adjustment of the dosage of the patient to be treated. Thepolymorph can be administered on a regimen of 1 to 4 times per day, suchas once or twice per day. This dosing regimen can be adjusted to providethe optimal therapeutic response.

Unit doses can be administered more than once a day, for example, 2, 3,4, 5 or 6 times a day. In various aspects, such unit doses can beadministered 1 or 2 times per day, so that the total dosage for a 70 kgadult human is in the range of 0.001 to about 15 mg per kg weight ofsubject per administration. In a further aspect, dosage is 0.01 to about1.5 mg per kg weight of subject per administration, and such therapy canextend for a number of weeks or months, and in some cases, years. Itwill be understood, however, that the specific dose level for anyparticular subject will depend on a variety of factors including theactivity of the specific polymorph employed; the age, body weight,general health, sex and diet of the individual being treated; the timeand route of administration; the rate of excretion; other drugs thathave previously been administered; and the severity of the particulardisease undergoing therapy, as is well understood by those of skill inthe area.

It can be necessary to use dosages outside these ranges in some cases aswill be apparent to those skilled in the art. Further, it is noted thatthe clinician or treating physician will know how and when to start,interrupt, adjust, or terminate therapy in conjunction with individualpatient response.

The present invention is further directed to a method for themanufacture of a medicament for treating disease relating to infectionby paramyxovirus in animals, including humans comprising combining oneor more disclosed polymorphs, products, or compositions with apharmaceutically acceptable carrier or diluent. Thus, in one aspect, theinvention relates to a method for manufacturing a medicament comprisingcombining at least one disclosed polymorph or at least one disclosedproduct with a pharmaceutically acceptable carrier or diluent.

The disclosed pharmaceutical compositions can further comprise othertherapeutically active polymorphs, which are usually applied in thetreatment of the above-mentioned pathological conditions.

In one aspect, the invention relates to pharmaceutical compositionscomprising the disclosed polymorphs. That is, a pharmaceuticalcomposition can be provided comprising a therapeutically effectiveamount of at least one disclosed polymorph or at least one product of adisclosed method and a pharmaceutically acceptable carrier.

In certain aspects, the disclosed pharmaceutical compositions comprisethe disclosed polymorphs as an active ingredient, a pharmaceuticallyacceptable carrier, and, optionally, other therapeutic ingredients oradjuvants. The instant compositions include those suitable for oral,inhaled, rectal, topical, and parenteral (including subcutaneous,intramuscular, and intravenous) administration, although the mostsuitable route in any given case will depend on the particular host, andnature and severity of the conditions for which the active ingredient isbeing administered. The pharmaceutical compositions can be convenientlypresented in unit dosage form and prepared by any of the methods wellknown in the art of pharmacy.

In various aspects, the invention also relates to a pharmaceuticalcomposition comprising a pharmaceutically acceptable carrier or diluentand, as active ingredient, a therapeutically effective amount of adisclosed polymorph, a product of a disclosed method of making, solvate,or a hydrate thereof. In a further aspect, a disclosed polymorph, aproduct of a disclosed method of making, solvate, hydrate thereof, maybe formulated into various pharmaceutical forms for administrationpurposes.

In practice, the polymorphs of this invention can be combined as theactive ingredient in intimate admixture with a pharmaceutical carrieraccording to conventional pharmaceutical compounding techniques. Thecarrier can take a wide variety of forms depending on the form ofpreparation desired for administration, e.g., oral, inhaled orparenteral (including intravenous). Thus, the pharmaceuticalcompositions of the present invention can be presented as discrete unitssuitable for oral administration such as capsules, cachets or tabletseach containing a predetermined amount of the active ingredient.Further, the compositions can be presented as a powder, as granules, asparticles, as a solution, as a suspension in an aqueous liquid, as anon-aqueous liquid, as an oil-in-water emulsion or as a water-in-oilliquid emulsion. The compositions may be formulated for administrationvia inhalant methods and devices. In addition to the common dosage formsset out above, the polymorphs of the invention, can also be administeredby controlled release means and/or delivery devices. The compositionscan be prepared by any of the methods of pharmacy. In general, suchmethods include a step of bringing into association the activeingredient with the carrier that constitutes one or more necessaryingredients. In general, the compositions are prepared by uniformly andintimately admixing the active ingredient with liquid carriers or finelydivided solid carriers or both. The product can then be convenientlyshaped into the desired presentation.

It is especially advantageous to formulate the aforementionedpharmaceutical compositions in unit dosage form for ease ofadministration and uniformity of dosage. Unit dosage form as used hereinrefers to physically discrete units suitable as unitary dosages, eachunit containing a predetermined quantity of active ingredient calculatedto produce the desired therapeutic effect in association with therequired pharmaceutical carrier. Examples of such unit dosage forms aretablets (including scored or coated tablets), capsules, pills, powderpackets, wafers, suppositories, injectable solutions, metered inhalants,or suspensions and the like, and segregated multiples thereof.

Thus, the pharmaceutical compositions of this invention can include apharmaceutically acceptable carrier and a polymorph as disclosed herein.The polymorphs of the invention can also be included in pharmaceuticalcompositions in combination with one or more other therapeuticallyactive compounds.

The pharmaceutical carrier employed can be, for example, a solid,liquid, or gas. Examples of solid carriers include lactose, terra alba,sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, andstearic acid. Examples of liquid carriers are sugar syrup, peanut oil,olive oil, and water. Examples of gaseous carriers include carbondioxide and nitrogen. It should be emphasized that the above-describedembodiments of the present device and process, particularly, and“preferred” embodiments, are merely possible examples of implementationsand merely set forth for a clear understanding of the principles of thedisclosure. All these and other such modifications and variations areintended to be included herein within the scope of this disclosure andprotected by the following claims. Therefore the scope of the disclosureis not intended to be limited except as indicated in the appendedclaims.

The following specific examples will illustrate the invention as itapplies to the methods of treatment using inhalers. It will beappreciated that other examples, including minor variations inprocedures will be apparent to those skilled in the art, and that theinvention is not limited to these specific illustrated Examples.

EXAMPLES Crystallization Process Development of P-2 and P-3 CrystalForms General

Experiments were carried out to define the crystallization space (inparticular, solvent composition and cooling rate) and proved that mostimportant process-relevant crystal forms P-2 hydrate, P-3 ethanolate andP-3 dihydrate can be produced in a controllable fashion.

Example 1

P-3 recrystallization/purification (example of a pilot batch): 540 mg ofMDT-637 (starting P-3 ethanolate material with purity approximately96.5%), was suspended in 13.5 mL of hot acetone at 55±5° C. Theresulting mixture was diluted with water (4.4 mL) at 55±5° C. resultingin a clear solution. The solution was added slowly to 18 mL of absoluteethanol with continuous stirring. The clear solution was allowed to stirwhile cooling naturally to room temperature and stirred forapproximately 3 hours. Nucleation was slow, but a heavy precipitateformed during the course of the room temperature stir. The solids werecollected by filtration and washed on the funnel with absolute ethanol(3×1 mL). The collected product vacuum dried at 55±5° C. affording 400mg (74%), HPLC: (99.3 A %), DSC: (onset 256.58° C./peak 262.70° C.), ¹HNMR: Consistent with structure with residual ethanol. PXRD: form P-3ethanolate obtained. Product was washed with water and showed conversioninto P-3 dihydrate according to the Karl Fischer (KF) analysis (6.8%)and absence of ethanol by ¹H NMR.

Example 2

P-2 hydrate (recrystallization/purification, demo batch): MDT-637 70.5 g(starting P-3 ethanolate material with purity approximately 96.5%) wassuspended in 1,763 mL of acetone at 55±5° C. The mixture was dilutedwith water (571 mL) while maintaining the temperature at 55±5° C. duringthe addition. The resulting mixture was allowed to stir at 55±5° C. forapproximately 15 minutes. The heat was removed and the mixture coolednaturally to room temperature and stirred overnight. The precipitatedsolids collected by filtration and washed on the funnel with a 1:1mixture of acetone/water (3×150 mL). The collected solids vacuum driedat 55±5° C. affording 55.7 grams (79%) product. ¹H NMR: consistent withstructure P-2 with no evidence of residual ethanol or acetone, HPLC:(99.4 A %), KF: (1.2%).

Example 3

Definition of the crystallization space for P-3 ethanolate crystal form(a): Starting P-3 pattern ethanolate was re-crystallized fromaqueous-acetone/ethanol (original volume of ethanol used/aq-acetone(0.82 v EtOH:1 v aq-acetone). Equilibrate at room temperature (R.T). for24 hours. Cool to 0-5° C. for at least 8 hrs. Material with pattern P-3was obtained.

Example 4

Definition of the crystallization space for P-3 ethanolate crystal formce (b): Starting P-3 pattern ethanolate was re-crystallized fromaqueous-acetone/ethanol (20% excess ethanol compared to Example 3).(1.15 v EtOH:1 v aq-acetone). Equilibrate at R.T. for 24 hours. Cool to0-5° C. for at least 8 hrs. Material with pattern P-3 was obtained.

Example 5

Definition of the crystallization space for P-3 ethanolate crystal form(c): Starting P-3 pattern ethanolate was re-crystallized fromaqueous-acetone/ethanol (20% less ethanol than in Example 3). (0.5 vEtOH:1 v aq-acetone). Equilibrate at R.T. for 24 hours. Cool to 0-5° C.for at least 8 hrs. Material with pattern P-3 was obtained.

Example 6

Definition of the crystallization space for P-3 ethanolate crystal form(d): Starting P-3 pattern ethanolate was re-crystallized fromaqueous-acetone/ethanol/water: (1 v aqeous-acetone:1.7 v (1:1ethanol/water). Equilibrate at R.T. for 24 hours. Cool to 0-5° C. for atleast 8 hrs. Material with pattern P-2 anhydrous form was obtained.

Example 7

Definition of the crystallization space for P-3 ethanolate crystal form(e): Starting P-3 pattern ethanolate was re-crystallized fromaqueous-acetone/ethanol (original volume of ethanol used/aq-acetone(0.82 v EtOH:1 v aq-acetone). The hot aq-acetone solution will be addedto cold (0-5° C.) ethanol and the resulting mixture further cooled to0-5° C. and equilibrated at 0-5° C. for approximately 8 hours. Partiallyamorphous material with pattern P-3 (low crystallinity) was obtained.

Example 8

Definition of the crystallization space for P-3 ethanolate crystal form(f): Starting P-3 pattern ethanolate was re-crystallized fromaqueous-acetone/ethanol (original volume of ethanol used/aq-acetone(0.82 v EtOH:1 v aq-acetone). The hot aq-acetone solution will be addedto hot (55±5° C.) ethanol and the resulting mixture slowly cooled to0-5° C. at a rate of ˜0.2° C./min. and equilibrated at 0-5° C. forapproximately 8 hours. Material with pattern P-3 was obtained. Thus theExamples 1-8 show that production of P-3 pattern ethanolate can becontrolled with sufficient amount of ethanol antisolvent (down to 33%v/v ethanol). However excess of water (Example 6) led to P-2 materialcrystallized. Also very rapid cooling crystallization may lead to lowcrystallinity product (Example 7).

Example 9

Equilibrium solubility of different API forms in aqueous ethanol: Thissolubility was determined for P-2 and P-3 crystal forms using slurryequilibration and solution concentration measurement with HPLC. Thefollowing solubilities are observed at room temperature in 75/25% v/vethanol/water: P-3 ethanolate: 239.9 μg/mL; P-2 hydrate: 416.5 μg/mL.These data indicate that P-3 ethanolate P-3 ethanolate is least soluble,and therefore more stable form than P-2 in ethanol/water mixture, As aresult, reproducible crystallization process for P-3 ethanolate can beachieved using a thermodynamically-controlled crystallization process.

Example 10

Precipitation of P-2 dihydrate from NMP solution: 594.7 mg of API wasdissolved in 20 mL of NMP solvent by stirring and slow heating to 60° C.The resulting stock solution was added to 300 mL pure water and left for14 hours. The precipitate was washed with water and dried at 40° C. 418mg of product was obtained which showed to be a partially amorphous P-2dihydrate form by PXRD analysis.

Example 11

Precipitation of P-2 anhydrous from DMF solution: 8% w/v of API in DMFsolution was prepared by stirring and slow heating to 60° C. 5 mL ofthis stock solution was added to 170 mL methanol antisolvent. Slowprecipitation was observed for 2 hours. The precipitate was washed withmethanol and dried at 40° C. 260 mg of product was obtained whichexhibited the structure consistent with P-2 anhydrous form by PXRDanalysis.

Precipitation of Other Forms and Compositions Example 12

Precipitation of P-4 material DMF solution: 300 mg of API was dissolvedin 5 mL of DMF solvent by stirring and slow heating to 60° C. Theresulting stock solution was placed in 50 mL open beaker to slowlyevaporate for 4 days at room temperature. The precipitate (166 mg) wascollected and analyzed using PXRD. The product was identified with P-4pattern and consisted presumably a DMF solvate form because this patternwas only repeated during evaporation, cooling or antisolventprecipitation with DMF.

Example 13

Precipitation of P-6 material from DMF solution: 5 mL of the 8% w/v APIstock solution in DMF was added to 75 mL of MTBE antisolvent. Fastprecipitation was observed. The precipitate was washed with MTBE anddried at 40° C. 264 mg of product was obtained which exhibited thestructure identified with P-6 pattern by PXRD analysis.

Example 14

Precipitation of P-7 material: 10 mL of 4% w/v API stock solution in NMPwas added to 75 mL of cyclohexane. No precipitation was observed forseveral hours and solvent phase separation occurred. Consequently, 100ml of water was added and the ternary mixture was stirred for 20 minutesand left to equilibrate for 14 hours. The precipitate was washed withwater and dried at 40° C. 380 mg of product was obtained which exhibitedthe structure identified with P-7 pattern by PXRD analysis.

Example 15

Precipitation of P-8 material: 5 mL of 8% w/v API stock solution in DMSOwas added to 100 mL of water. Fast precipitation was observed. Theprecipitate was washed with water and dried at 40° C. 308 mg of productwas obtained which exhibited the structure identified with P-8 patternby PXRD analysis.

Example 16

Precipitation of amorphous material: 100 mg of API was suspended in 3 mLof hot acetone at 55 f 5° C. The resulting mixture was diluted withwater (0.7 mL) at 55±5° C. resulting in a clear solution. The solutionwas cast on a glass plate heated at 60° C. 60 mg of the product obtainedexhibited no distinct diffraction peaks when analyzed with PXRD and wasidentified with amorphous structure.

Example 17

Co-precipitation of API with excipient: 100 mg of API was suspended in13.5 mL of hot acetone at 55±5° C. The resulting mixture was dilutedwith water (4.4 mL) at 55±5° C. resulting in a clear solution. 500 mg ofhydroxypropyl methylcellulose acetate succinate (HPMCAS) was dissolvedin the same solution. The solution was cast on a glass plate heated at60° C. 420 mg of the product obtained exhibited no distinct APIdiffraction peaks when analyzed with PXRD and was identified withamorphous dispersion of API in the HPMCAS polymer matrix.

Equilibrium Solubility and Thermodynamic Stability of P2 and P-3 CrystalForms Example 18

FIG. 23 shows the results of solubility study of drug concentration instirred suspensions at 25° C. as a function of time for P-2, P-3 formsand their mixture in acetonitrile-water 50/50 v/v solution. It indicatesabout 40% lower solubility of P-2 compared to P-3 at 25° C. A mixture offorms: P-3 (approximately 75% w/w) and P-2 (approximately 25% w/w), hasshown an intermittent solubility. As a function of time, there was aclear reduction of solubility for both P-3 and P-3+P-2 materials. Thesedata indicate lower solubility, greater thermodynamic stability of P-2and conversion of P-3 form into a less soluble P-2 form at a giventemperature.

Example 19

Competitive slurry stability studies over 10 days between P-2 and P-3crystal forms in 50/50 acetonitrile/water mixture at 25° C. showed thatP-3 converts to P-2 as determined using PXRD quantitative phase analysis(based on assessment of multiple peaks ratio). The composition of slurryfor P-2 hydrate changed from 50% (day 0) to 77% (4 days) to 85% (10days). Correspondent solubilities in this solvent system were found tobe 190 μg/mL (P-2) and 310 μg/mL (P-3). Therefore P-2 hydrate is morethermodynamically stable at given temperature.

Example 20

An equal powder mixture of six known phases (P-2, P-3, P-4, P-6, P-7 andP-8), 50 mg each were stirred in 75/25 acetone-water solvent (50 mL) at25° C. over 12 days. PXRD analysis showed that all forms convertedconvert to P-2 hydrate, which is shown to be the most thermodynamicallystable form at these conditions.

Example 21

Simulated lung fluid was prepared according to a method described byMoss et al. (Health Physics 1979, v36, 447-448). The composition ofsimulated lung fluid (pH 7.4) is showed in TABLE 12.

TABLE 12 Concentration Chemical (g/L) Magnesium chloride MgCl₂•6H₂O0.2033 Sodium chloride NaCl 6.0193 Potassium chloride KCl 0.2982 Sodiumhydrogen phosphate Na₂HPO₄•12H₂O 0.3582 Sodium sulphate Na₂SO₄ 0.0710Calcium chloride CaCl₂•2H₂O 0.3676 Sodium acitate CH₃COONa•3H₂O 0.9526Sodium bicarbonate NaHCO₂ 2.6043 Sodium citrate Na₃H

C

O

•2H₂O 0.0970

indicates data missing or illegible when filed

A 0.1% DPPC in water solution was prepared according to a procedurepublished by Marques et al. (Dissolution Technologies, August 2011). TheSLF solution modified with 0.02% DPPC was prepared by diluting the 0.1%DPPC solution with SLF. Refer to NB1181P59-60 for the details on thepreparation procedure of SLF and SLF modified with 0.02% DPPC. Two typesof DPPC were used in the study. The hydrogenated DPPC was supplied byLipoid LLC (USA), while the non-hydrogenated DPPC was supplied bySigma-Aldrich (catalogue # P0763-1G). SLF modified with 0.02% DPPCappeared to be a cloudy/milky suspension, which indicated the formationof micelles and/or emulsions. In order to segregate the solid drugsubstance from the cloudy solution, a dialysis tube (Sigma-Aldrichcatalogue # D9277-100FT) was used. A saturated solution of the drugsubstance in SLF modified with 0.02% DPPC was prepared and carefullytransferred into the dialysis tube (˜10 mL in volume). The tube was thenput in a container filled with about 100 mL of SLF modified with 0.02%hydrogenated DPPC or 0.02% DPPC. A working bottle was used as thecontainer in the study for SLF with 0.02% hydrogenated DPPC. However, itwas determined later that the dialysis tube had better lay-out in acrystallization dish with less turns/kinks. Therefore, a crystallizationdish was used as the container in the study of SLF with 0.02% DPPC. Foreach lot of the drug substance, three samples were set up and placedinto a glove box (supplied by Coy Labs) maintained at 37° C. The sampleswere put on a rotator, which was turned on afterwards at relatively lowspeed. Samples were taken at different time points for HPLC analysis. Asample preparation procedure for HPLC analysis was developed for samplesin SLF modified with 0.02% hydrogenated DPPC or 0.02% DPPC. The samplesolution was diluted by 1:3 ratio with ethanol. About one hour after thedilution, it was filtered for HPLC analysis. Glass volumetric pipetteswere used and the problem was resolved. The results are shown in FIG.18.

Chemical and Formulation Stability Example 22

Forced degradation experiments consisted of placing solid samples of P-2hydrate and P-3 dihydrate materials in sealed glass bottles at 120° C.for 7 days. The following results were obtained: (a) P-3 sample: APIdecreased from 98.7% to 93.3%; 6 peaks in control to 16 peaks instressed sample. (b) P-2 sample: MDT-637 decreased from 99.2% to 99.1%;4 peaks in control to 5 peaks in stressed sample. The data are shown inTABLE 13. Batche P224-135-2 corresponds to P-2 hydrate and 224-163-1corresponds to P-3 dihydrate. Thus P-2 form showed to be a more stableform against accelerated solid-state chemical degradation than P-3dihydrate.

TABLE 13 Impurity/ Degradation MDT- Product → A B C D E F G H 637 I J KL RT 1.18 2.08 2.79 3.16 9.29 10.51 12.23 13.79 16.05 17.44 18.71 19.1120.07 (minutes) → Relative 0.07 0.13 0.17 0.20 0.58  0.66  0.76  0.86 NA 1.09  1.17  1.19  1.25 RT → Sample Area % Name P224-163-1 0.09 — — — —— —  0.54 98.70  0.24  0.05  0.16  0.06 CTRL P224-163-1 0.12 3.60 0.170.48 0.28  0.08  0.05  0.43 93.32  0.17 —  0.13  0.06 120C/7D P224-135-20.06 — — — — — —  0.36 99.22  0.09 —  0.16 — CTRL P224-135-2 0.07 — — —— — —  0.40 99.07  0.07 —  0.16 — 120C/7D Impurity/ Total DegradationImp/ Grand Product → M N O P Q Deg Total RT (minutes) → 20.93 21.3522.84 23.68 26.02 NA NA Relative RT →  1.30  1.33  1.42  1.48  1.62 NANA Sample Name Area % P224-163-1 CTRL — — — — — 1.16 99.9 P224-163-1120C/7D  0.09  0.07  0.37  0.36  0.05 6.49 99.8 P224-135-2 CTRL — — — —— 0.67 99.9 P224-135-2 120C/7D — — —  0.06 — 0.76 99.8

Example 23

The micronized, P-3 dihydrate material was stored in 1.85 mL amberbottles with Teflon lined caps. The 25° C./60% RH and 40° C./75% RHsimulated storage and worst-case storage conditions correspondingly.Data through 12 months for API storage in glass bottles indicated thatno change in form was observed according to both DSC and PXRD analyses.

Example 24

P-3 dihydrate and P-2 hydrate materials were placed in open wide glassbottles at 40° C./75% RH for 4 weeks. The results indicated no change ofform by both DSC and PXRD analyses. Acceptable chemical stability wasobserved for both forms as shown in TABLE 14.

TABLE 14 Attribute P-2 P-3 Assay 98.8% 96.7% Rel Subs 0.68% 0.67% Water 3.5%  5.9%

Example 25

Formulation of the P-3 dihydrate form of the API with lactose (6.3% w/w)was placed on stability 40° C./75% RH in sealed glass bottles for 39weeks. After samples removal, the lactose was removed by dissolution inpurified water. The reference formulation was also subjected to the samelactose extraction procedure to eliminate the possibility for formconversion in water. The stability samples indicated no change ofcrystal form.

Mieronization and Formulation

Example 26

Four lots of MDT-637 API were micronized using a 2-inch spiral jet mill.The approximate amounts of material available from each of theexperiments and the initial particle size distributions (PSDs) of theunmilled materials are list in Table 15.

TABLE 15 Quantity X₁₀ X₅₀ X₉₀ Lot (g) (μm)* (μm)* (μm)* XRPD PatternP218-75-4 50 0.81 2.77 16.37 P-3/P-2 (predominantly P-3) P224-135-2 501.28 10.35 77.40 P-2 P224-131-2 50 1.36 10.61 78.99 P-2 P224-177-1 60.92 3.67 19.89 P-3

The resulting particle size distribution (PSD) percentiles are shown inTABLE 16 for micronized P-2 hydrate, P-3 dihydrate and a physicalmixture materials.

TABLE 16 Mill- ing Pres- X₁₀ X₅₀ X₉₀ Sample sure Lot (μm) (μm) (μm)Number (psi) P218-75-4 First Pass, 0.49 1.16 2.45 I-1 120 50 g Second0.47 1.10 2.20 I-2 120 Pass, 25 g P224-135-2 25 g (initial) 0.59 1.674.62 I-3 80 25 g (final) 0.60 1.73 5.29 I-4 80 P224-131-2 25 g(initial), 0.65 1.96 5.58 I-5 80 First Pass 25 g (initial), 0.62 1.775.16 I-7, re- 20 Second Pass pass 25 g (final) 0.62 1.80 5.08 I-6 110P224-177-1 1 pass 0.53 1.19 2.49 I-8 80

It was noticed that at higher milling pressure the P-2 hydrate materialyielded larger particles compared to P-3 dihydrate material which wasattributed to higher hardness of the P-2 hydrate compound as bothmaterials show a similar acicular particle shape. Micronization of bothforms could be sufficiently controlled to produce PSD in a desiredrespiratory size range (e.g. D₅₀ between 1-2 μm and D₉₀<5 μm). Inaddition, micronization did not affect the crystallinity, phase purityof both P-3 dihydrate and P-2 hydrate forms.

Example 27

Safety and Tolerability of MDT-637 P3

The purpose of the following studies was to assess the safety,tolerability, local and systemic pharmacokinetics of single and multipledoses of MDT-637 P3 in healthy volunteers and asthmatics.

For Phase 1 studies, an inhaler such as the MicroDose Inhaler (anactive, electronic DPI) was developed to synchronize drug aerosolizationwith adult tidal breathing, and adapted to fit a valved aerosol mask(FIG. 24). In summary, the MicroDose Inhaler operation comprises thedrug powder contained in a protective blister until delivery. Theblister is pierced externally and then placed in contact with apiezoelectric vibrator within the MicroDose Inhaler. When the patientinhales, an airflow sensor automatically turns on the piezo whichprovides energy to deaggregate the particles of powder and aerosolizesthem into the inhalation airstream, providing synchronized delivery. TheMicroDose Inhaler was triggered by pressure drop and programmed toproduce a series of brief aerosol bursts of predetermined durationwithin the range of 0.1 to 2.0 seconds, preferably 0.1 seconds, earlyduring each inhalation cycle and for 16 consecutive breaths. The studieswere conducted as follows:

Study 1: A Randomized, Double-blind, Placebo-controlled, SingleAscending Dose (SAD) Study to Assess the Safety, Tolerability andPharmacokinetics of Inhaled MDT-637 in Healthy Volunteers (SAD Trial)

Study Objective:

The objective of Study 1 was to determine the range of inhaled doses ofMDT-637 P3 that are safe and well-tolerated including effects on “ForcedExpiratory Volume in the First Second” (FEV₁). An additional objectivewas to determine the rate and extent of systemic absorption of MDT-637P3 and trough nasal wash MDT-637 P3 drug concentrations after a nasalinhalation.

Study Design:

Single-center, double-blind, randomized, placebo-controlled, sequentialgroup.

Each subject received (Once Daily or Three Times Daily (TID) over asingle day) nasally inhaled dose(s) of MDT-637 P3 (or placebo) listedbelow. The study consisted of 3 visits: Visit 1 (Screening), Visit 2(Dosing) and Visit 3 (Follow up); subjects furloughed from the clinicbetween Visits 2 and 3. Subjects who met all of the inclusion criteriaand who meet none of the exclusion criteria (Table 17, FIG. 27) at Visit1 were eligible to return for Visit 2 within 14 days of screening. Thesubject was admitted to the study center the day prior to dosing, wasobserved at the study center for at least 24 hours after dosing, andreturned for Visit 3, 7 days after dosing.

Groups of 8 subjects were enrolled into each of the following cohortsrandomized 3:1 (active:placebo):

Treatment (expressed as target emitted dose exiting Treatment Cohort thefacemask) 1 MDT-637, 8 mcg Once Daily or placebo 2 MDT-637, 33 mcg OnceDaily or placebo 3 MDT-637, 33 mcg TID or placebo 4 MDT-637, 66 mcg OnceDaily or placebo 5 MDT-637, 66 mcg TID or placebo 6 MDT-637, 132 mcg 3times daily or placebo

Study Drug and Formulation

Placebo: Inhalation grade lactose blend (100% RESPITOSE® ML003(Princeton, N.J., USA))

Active: Micronized MDT-637 P3 formulated^(w/w) as a dry powder withinhalation grade lactose (ML003)

Low strength formulation: 0.63%^(w/w) MDT-637 in RESPITOSE® ML003 Highstrength formulation: 6.3%^(w/w) MDT-637 in RESPITOSE® ML003

Dose and Route of Administration

The doses of MDT-637 P3 were determined based on the no adverse effectlevel (NOAEL) in GLP toxicology studies in two species, as well as onthe basis of previous clinical experience with a previous inhaledsolution formulation (referred to as VP14637 Drug Product). Both activeand matched placebo were administered via face mask inhalation from aproprietary delivery system (MicroDose inhaler).

At each dose level, 6 subjects received MDT-637 P3 and 2 subjectsreceived matched placebo. Based on safety and tolerability results fromeach cohort, the Principal Investigator, Medical Monitor and MDT jointlydecided whether to proceed to the next dose level based uponprotocol-specified dose escalation rules. At least 7 days (between Visit2 dosing days in subsequent dose groups) separated successive dosingcohorts. A second cohort was available to be studied at any dose levelif needed to confirm findings from the initial cohort at that dose.

Single doses were administered in the morning. TID doses will beadministered at 6 hour intervals, with the 1st of the 3 dosesadministered in the morning.

Assessments

Primary Endpoints

Safety and tolerability, were assessed by:

Vital signs at Visits 1, 2 and 3

Physical examination at Visit 1, 2 and 3

Routine laboratory tests (hematology, clinical chemistry,

urinalysis) at Visit 1 (screening), Visit 2 and Visit 3 (discharge)

ECGat Visits 1, 2 and 3

Spirometry (FEV1) assessed at Visit 1 (screening) and Visit 2 i.e.,comparing pre and post dosing

Adverse event assessment from administration of study drug

until subjects were discharged from the study

Secondary Endpoints

Pharmacokinetic endpoints (Cmax, Tmax, AUC, T1/2) and doseproportionality were assessed based on a limited number of plasmasamples from Visit 2

Analysis

Demographic and baseline information was presented and summarized bytreatment sequence and across the entire group. Subjects who wereenrolled in the study and received study treatment could have beenreplaced (at the discretion of the Sponsor) if they discontinued priorto the completion of the study. Subjects were not replaced if they werediscontinued from the study secondary to an adverse event/adverseexperience (AE) unless the AE could be determined to be unrelated totreatment.

Safety and tolerability were evaluated based on the results of physicalexamination, electrocardiogram (ECG) (with QTc intervals), laboratorytests (urinalysis, hematology, chemistry), spirometry and adverse eventassessments following study drug administration.

Pharmacokinetic variables (Cmax, Tmax, AUC, T1/2) were evaluated basedon blood samples drawn at Visit 2.

Study 2: A Randomized, Double-blind, Placebo-controlled Study to Assessthe Safety, Tolerability and Pharmacokinetics of Inhaled MDT-637 (P3)Administered to Healthy Volunteers for 10 Days (Multiple Ascending Dose:MAD Trial)

Study Objective:

The objective of Study 2 was to assess the tolerability and safety of arange of repeated inhaled doses of MDT-637 P3 that had been developedfor patients with RSV infection. A second objective of Study 2 was todetermine the rate and extent of systemic absorption of MDT-637 P3 andnasal mucosal MDT-637 P3 drug concentrations across 10 days of dosing.

Study Design:

Single-center, double-blind, randomized, placebo-controlled, sequentialgroup.

Subjects were enrolled in cohorts of 12 subjects. Within each cohort, 9subjects received nasally inhaled dose(s) of MDT-637 P3 and 3 subjectsreceived matching placebo. The study consisted of 3 visits: Visit 1(Screening), Visit 2 (Dosing), Visit 3 (Follow up) and subjects remainedat the study site for pharmacokinetic (PK) sampling, spirometry, andsafety assessments. Subjects who met all of the inclusion criteria andwho meet none of the exclusion criteria (Table 18, FIG. 28) wereeligible for this study. Day 1 was the first day of dosing and Day 10was the final day of dosing, following which subjects remained at thestudy site for 24 hours following the last dose for PK sampling andspirometry. Visit 3 was a follow-up visit, 7 to 10 days after the lastdose. Cohorts were dosed as follows:

Treatment Treatment (expressed as target emitted dose exiting Cohort thefacemask) 1 MDT-637 P3, 66 mcg twice daily (BID) or placebo 2 MDT-637P3, 66 mcg three times daily (TID) or placebo 3 MDT-637 P3, 132 mcg TIDor placebo

Study Drug and Formulation

Placebo: Inhalation grade lactose processed in identical manner toactive clinical blend (100% RESPITOSE® ML003)

Active: Micronized MDT-637 P3 formulated at 6.3%^(w/w) as a dry powderwith inhalation grade lactose (RESPITOSE® ML003)

Dose and Route of Administration

The doses of MDT-637 were determined based on the NOAEL in GLPtoxicology studies, as well as on the basis of previous clinicalexperience in Study 1 (above). Both active and matched placebo wasadministered via facemask using a MicroDose inhaler device.

At each dose level, 9 subjects received MDT-637 P3 and 3 subjectsreceived matching placebo. Based on safety and tolerability results fromeach cohort, the Principal Investigator, Medical Monitor and the Sponsor(MDT) jointly decided whether to proceed to the next dose level basedupon protocol-specified dose escalation rules. Dose escalation fromCohort 5 (66 mcg TID) to Cohort 6 in Study 1 was required for theinitiation of Cohort 1 in this study. At least 7 days separated the lastdose (i.e. Day 10) for a given cohort from the initiation of dosing inthe subsequent dosing cohort, following review of all safety data and adecision made to dose escalate. A second cohort may have been studied atany dose level if needed to confirm findings from the initial cohort atthat dose.

The first doses for twice daily (BID) and three times daily (TID) wereadministered in the morning, starting at approximately 0700 hours oneach day of dosing. BID doses were administered at 12 hour intervals andTID doses were administered at 6 hour intervals each dose day.

Assessments

Primary Endpoints

Safety and tolerability, was assessed by:

Vital signs (blood pressure, heart rate, temperature and respiratory

rate) and pulse oximetry at Visits 2 and 3

Physical examination at Visits 2 and 3

Routine laboratory tests (hematology, clinical chemistry, urinalysis)

at Visits 2 and 3

ECG at Visits 2 and 3

Spirometry at Visits 2 and 3

Adverse event assessment from administration of study drug until

subjects are discharged from the study

Secondary Endpoints

Pharmacokinetic endpoints (Cmax, Tmax, AUC, T1/2) and doseproportionality was assessed based on a limited number of plasma samplesfollowing the first dose on Day 1 and last dose on Day 10

Trough plasma samples for MDT-637 P3 concentration was obtained within30 minutes prior to the first daily dose on Days 2, 5 and 10

MDT-637 concentrations determined from nasal wash samples prior to and15 minutes post dose on Day 6 and 24 hours post last dose (Day 11)

Analysis

Demographic and baseline information was presented and summarized bytreatment sequence and across the entire group.

Subjects who were enrolled in the study and received study treatment mayhave been replaced (at the discretion of the Sponsor) if theydiscontinued prior to the completion of the study. Subjects were notreplaced if they were discontinued from the study secondary to an AEunless the AE was determined to be unrelated to treatment.

Safety and tolerability were evaluated based on the results of physicalexamination, ECG, laboratory tests (urinalysis, hematology, chemistry),spirometry and adverse event assessments following study drugadministration.

Pharmacokinetic variables (Cmax, Tmax, AUC, T1/2) were evaluated basedon blood samples drawn following first dose (Days 1-2) and last dose(Days 10-11).

Study 3: A Double-Blind, Randomized, 3 Period Crossover, SingleAscending Dose Study to Assess the Safety and Tolerability of InhaledMDT-637 P3 in Subjects with Intermittent or Mild-to-Moderate PersistentAsthma (Asthma Trial)

Study Objective:

The primary objective of Study 3 was to assess the safety andtolerability of MDT-637 P3 when inhaled by subjects with Intermittent,or Mild-to-Moderate Persistent, Asthma. A secondary objective of Study 3was to determine the rate and extent of systemic absorption of MDT-637and trough nasal wash MDT-637 drug concentrations following dry powderinhalation dosing.

Study Design:

Single-center, double-blind, randomized, 3 period crossover, ascendingdose.

Each subject was dosed on 3 separate days during the study. Each subjectreceived an initial dose of placebo (Period 1), followed by randomallocation at Period 2 to a single dose of nasally inhaled MDT-637 P3 ordouble blind placebo. After an observation and furlough period, subjectsreturned for Period 3 to receive three doses of study drug (MDT-637 P3or placebo) over a single day as listed below. The study consisted of 5visits: Visit 1 (Screening), Visits 2, 3 and 4 (Dosing), and Visit 5(Follow up); subjects were furloughed from the clinic for 6 up to 14days between Visits 3 and 4 and 7-8 days between Visit 4 and 5. Subjectswho met all of the inclusion criteria and none of the exclusion criteria(Table 19, FIG. 29) at Visit 1 were eligible to return for Visit 2within 42 days of screening. Subjects were admitted to the study centertwice, once for Visit 2 and again for Visit 4. Visit 5, was a safetyfollow up (outpatient) visit.

A single group of approximately 10 subjects was enrolled into the study.

Treatment (expressed as target emitted dose exiting Visit the facemask)1 Screening 2 Placebo 3 Double-blind, randomized MDT-637, 66 mcg orplacebo Once Daily 4 Double-blind, randomized MDT-637, 132 mcg orplacebo TID 5 Follow-Up visit

Study Drug and Formulation

Placebo: Inhalation grade lactose processed in an identical manner toactive clinical blend (100% RESPITOSE® ML003)

Active: Micronized MDT-637 P3 formulated^(w/w) as a dry powder withinhalation grade lactose (ML003)

Dose and Route of Administration

The doses of MDT-637 P3 were determined based on previous clinicalexperience in Study 1. Both placebo and active were administered as adry powder for nasal inhalation via a face mask using a MicroDoseInhaler device. At Visit 2, all 10 patients were dosed with placebo OnceDaily. At Visit 3, subjects were randomly allocated so that 8 subjectswere dosed with MDT-637 P3 Once Daily 66 mcg and 2 subjects receiveddouble blind placebo. At Visit 4, subjects were again randomly allocatedso that 8 subjects were dosed with MDT-637 TID 132 mcg and 2 subjectsreceived double blind placebo.

Based on safety and tolerability results from each visit in Study 3 andsafety data from Study 1 and Study 2, the Principal Investigator,Medical Monitor and Sponsor (MicroDose Therapeutx, MDTx) could jointlydecide whether to proceed to the next dose administration based uponprotocol-specified dose escalation rules. At least 7 days (between Visit3 and Visit 4) separated successive dosing cohorts. A second cohortcould have been studied at any dose level if needed to confirm findingsat that dose. Single doses were administered in the morning, starting atapproximately 0700 hours. TID doses were administered at 6 hr intervals,with the first of the 3 doses administered at approximately the sametime in the morning as the Once Daily doses are administered.

Assessments

Primary Endpoints

Safety and tolerability, was assessed by:

Adverse event assessment from administration of study drug until

discharge from the study

Vital signs at Visits 2, 3, 4 and 5

Physical examination at Visits 2, 4 and 5

Routine laboratory tests (hematology, clinical chemistry, urinalysis) atVisits 2, 3, 4 and 5

ECG at Visits 2, 3, 4 and 5

Spirometry (FEV₁) at Visit 3 and Visit 4.

Secondary Endpoints

Pharmacokinetic endpoints (Cmax, Tmax, AUC, T1/2) were assessed based ona limited number of plasma samples from Visit 4

Trough MDT-637 P3 concentrations determined from nasal wash

Analysis

Demographic and baseline information was presented and summarized bytreatment and across the entire group.

Subjects who were enrolled in the study and received study treatment butwithdrew before study completion could have been replaced at thediscretion of the Sponsor. Subjects were not replaced if theydiscontinued from the study secondary to an AE unless the AE could bedetermined to be unrelated to treatment.

Safety and tolerability were evaluated based on the results of adverseevent assessments, physical examination, vital signs, spirometry, ECG(with QTc intervals), and laboratory tests (urinalysis, hematology,chemistry) following study drug administration.

Pharmacokinetic variables (Cmax, Tmax, AUC, T1/2) were evaluated basedon blood samples drawn at Visit 4. Nasal trough concentrations at Visit4 were listed.

Results and Discussion Study 1: Single Ascending Dose Study

Ascending doses tested from 2 mcg once daily to 132 mcg three timesdaily in 35 volunteers

MDT-637 P3 was safe and well tolerated at doses up to 132 mcg TID

Overall, 4 subjects (11.4%) reported experiencing 4 TEAEs during thestudy (No significant changes in pulmonary function or reports ofpulmonary AEs)

Mild dizziness was sole treatment emergent adverse event consideredpossibly related by PI

No clinically significant changes in labs, vital signs, ECGs, physicalexams

Nasal wash levels at 24 hours post dose were >MDT-637 P3's IC₅₀ for RSV

Consistent and low PK exposure (Mean C_(max) of 33.7±4.6 picograms/mL(132 mcg TID cohort)

Study 2 Multiple Ascending Dose Study

Ascending doses tested up to 132 mcg three times daily for 10 days in 38volunteers

MDT-637 P3 was safe and well tolerated at doses up to 132 mcg TID for 10days

No significant changes in pulmonary function or reports of pulmonary AEs(FIG. 25)

25 subjects reported no treatment emergent adverse events (TEAEs)

The remaining 13 subjects reported a total of 20 TEAEs overall, allconsidered mild

5 TEAEs were considered related to study treatment: eye twitching(possibly), dry throat (related), headache (possibly, placebo), sorethroat (possibly) and lethargy (possibly). No action was taken on any ofthe events and all reported as ‘resolved’

No clinically significant changes in labs, vital signs, ECGs, physicalexams

Nasal wash levels at pre/post-dose and 24 hours post dose were >MDT-637P3 IC₅₀ for RSV

Consistent and low PK exposure (FIG. 26), with minimal accumulation.

Day 10 mean C_(max) ˜50 picograms/mL for 132 mcg TID

AUC₀₋₂₄ was 1.16 ng·h/mL)

CVs in the 15-30% range (excellent consistency)

Study 3: Single Ascending Dose in Asthmatics

Ascending doses tested from 66 mcg once daily to 132 mcg TID in n=10subjects to rule out repeat dose irritancy in patients with moresensitive airways

MDT-637 P3 was safe and well tolerated in asthmatic patients

No clinically significant changes in pulmonary function or reports ofpulmonary AEs

No clinically significant changes in labs, vital signs, ECGs, physicalexams

8/10 subjects reported no adverse events

The remaining 2/10 subjects reported (1) unrelated menstrual migrainesymptoms during the placebo phase and (2) moderate headache and mildnausea considered possibly related.

Pharmacokinetics were comparable between the healthy and asthmaticpopulation

CONCLUSIONS

The above Studies demonstrate that MDT-637 P3 is safe and well toleratedin three studies at doses up to 132 mcg TID for 10 days.

MDT-637 P3 exhibited highly consistent and low systemic exposure, whichis considered ideal for pediatric dosing. Importantly, no seriousadverse events were observed. Nasal levels of MDT-637 P3 quantified 15min, 6 hr and 24 hr post dose were >RSV IC₅₀.

Finally, no pulmonary adverse events or changes in pulmonary functionwere observed and no clinically significant changes in ECG, laboratoryvalues, vital signs or physical exam were observed either.

It will be appreciated by those skilled in the art that the presentinvention can be embodied in other specific forms without departing fromthe spirit and essential characteristics thereof. The presentlydisclosed embodiments are therefore considered in all respects to beillustrative and not restricted. The scope of the invention is indicatedby the appended claims rather than foregoing description and all changesthat come within the meaning and range and equivalence thereof areintended to be embraced therein.

1. A composition comprising at least one polymorph of MDT-637 selectedfrom the group consisting of: P-2 hydrate crystal form havingcharacteristic powder X-ray diffraction peaks at 2θ values of 4.83°,8.42°, 9.61°, 11.83°, 14.60°, 16.94°, 19.25°, 20.46°, 24.09°, 24.85°,27.76° and 28.56°; P-3 monohydrate crystal form having characteristicpowder X-ray diffraction peaks at 2θ values of 8.25°, 9.55°, 11.66°,14.30°, 18.03°, 19.70°, 20.29°, 24.55°, 25.29°, 26.31° and 27.26°; P-3dihydrate crystal form having characteristic powder X-ray diffractionpeaks at 2θ values of 7.03°, 8.16°, double peak at 9.47° and 9.75°,11.60°, 14.24°, 17.95°, 19.64°, 20.26°, 24.52°, 26.24° and 27.25°; P-3ethanolate crystal form having characteristic powder X-ray diffractionpeaks at 2θ values of 8.25, 9.54, 9.85, 11.69, 14.32, 15.99, 18.06,19.73, 20.33, 24.54, and 26.27; P-3 monohydrate crystal form havingcharacteristic powder X-ray diffraction peaks at 2θ values of 8.25°,9.55, 11.66, 14.30, 18.03, 19.70, 20.29, 24.55, 25.29, 26.31 and 27.26;P-3 anhydrous crystal form having characteristic powder X-raydiffraction peaks at 2θ values of 8.27°, 9.54°, 11.68°, 14.29°, 18.07°,19.79°, 20.30°, 24.54°, 25.27°, 26.36°, and 27.25°; P-2 anhydrouscrystal form having characteristic powder X-ray diffraction peaks at 2θvalues of 8.58°, 10.64°, 12.58°, 14.51°, 15.89°, 17.20°, 21.06°, 21.41°,22.37°, 25.43° and 27.62°; P-4 crystal form having characteristic powderX-ray diffraction peaks at 2θ values of 4.31°, 7.99°, 9.37°, 11.02°,13.04°, 13.43°, 14.18°, 16.13°, 16.70°, 17.08°, 17.42° and 17.92°; P-6crystal form having characteristic powder X-ray diffraction peaks at 2θvalues of double peak at 3.43° and 3.89°, 6.89°, double peak at 7.87°and 8.25°, 10.88°, 13.06°, 13.81°, 16.12°, 17.38° and 18.51°; P-7crystal form having characteristic powder X-ray diffraction peaks at 2θvalues of 5.18°, 6.59°, 7.70°, double peak at 10.02° and 10.62°, doublepeak at 12.18° and 12.50°, 15.16°, double peak at 15.88° and 16.4°,double peak at 17.20° and 17.42°, and double peak at 18.07° and 18.25°;P-8 crystal form having characteristic powder X-ray diffraction peaks at2θ values of 4.15°, 7.85°, 9.33°, 14.29°, triple peak at 15.84°, 16.68°and 17.14°, 18.53°, 20.27°, 23.90°, 24.80° and 27.39°; and combinationsthereof.
 2. The composition of claim 1, wherein the compositioncomprises P-2 hydrate crystal form having characteristic powder X-raydiffraction peaks at 2θ values of 4.83°, 8.42°, 9.61°, 11.83°, 14.60°,16.94°, 19.25°, 20.46°, 24.09°, 24.85°, 27.76° and 28.56°.
 3. Thecomposition of claim 1, wherein the composition comprises P-3monohydrate crystal form having characteristic powder X-ray diffractionpeaks at 2θ values of 8.25°, 9.55°, 11.66°, 14.30°, 18.03°, 19.70°,20.29°, 24.55°, 25.29°, 26.31° and 27.26°.
 4. The composition of claim1, wherein the composition comprises P-3 dihydrate crystal form havingcharacteristic powder X-ray diffraction peaks at 2θ values of 7.03°,8.16°, double peak at 9.47° and 9.75°, 11.60°, 14.24°, 17.95°, 19.64°,20.26°, 24.52°, 26.24° and 27.25°.
 5. The composition of claim 1,wherein the composition comprises P-3 ethanolate crystal form havingcharacteristic powder X-ray diffraction peaks at 2θ values of 8.25°,9.54°, 9.85°, 11.69°, 14.32°, 15.99°, 18.06°, 19.73°, 20.33°, 24.54°,and 26.27°;
 6. The composition of claim 1, wherein the compositioncomprises P-3 anhydrous crystal form having characteristic powder X-raydiffraction peaks at 2θ values of 8.27°, 9.54°, 11.68°, 14.29°, 18.07°,19.79°, 20.30°, 24.54°, 25.27°, 26.36°, and 27.25°.
 7. The compositionof claim 1, wherein the composition comprises P-2 anhydrous crystal formhaving characteristic powder X-ray diffraction peaks at 2θ values of8.58°, 10.64°, 12.58°, 14.51°, 15.89°, 17.20°, 21.06°, 21.41°, 22.37°,25.43° and 27.62°.
 8. The composition of claim 1, wherein thecomposition comprises P-4 crystal form having characteristic powderX-ray diffraction peaks at 2θ values of 4.31°, 7.99°, 9.37°, 11.02°,13.04°, 13.43°, 14.18°, 16.13°, 16.70°, 17.08°, 17.42° and 17.92°. 9.The composition of claim 1, wherein the composition comprises P-6crystal form having characteristic powder X-ray diffraction peaks at 2θvalues of double peak at 3.43° and 3.89°, 6.89°, double peak at 7.87°and 8.25°, 10.88°, 13.06°, 13.81°, 16.12°, 17.38° and 18.51°.
 10. Thecomposition of claim 1, wherein the composition comprises P-7 crystalform having characteristic powder X-ray diffraction peaks at 2θ valuesof 5.18°, 6.59°, 7.70°, double peak at 10.02° and 10.62°, double peak at12.18° and 12.50°, 15.16°, double peak at 15.88° and 16.4°, double peakat 17.20° and 17.42°, and double peak at 18.07° and 18.25°.
 11. Thecomposition of claim 1, wherein the composition comprises P-8 crystalform having characteristic powder X-ray diffraction peaks at 2θ valuesof 4.15°, 7.85°, 9.33°, 14.29°, triple peak at 15.84°, 16.68° and17.14°, 18.53°, 20.27°, 23.90°, 24.80° and 27.39°.
 12. The compositionof claim 1, additionally comprising one or more pharmaceuticallyacceptable carriers.
 13. The composition of claim 1, wherein the atleast one polymorph of MDT-637 is dispersed in at least onepharmaceutical excipient to provide a solid composite.
 14. A compositioncomprising at least one polymorph of MDT-637 selected from the groupconsisting of: P-2 hydrate crystal form characterized by a powder X-raydiffraction pattern as shown in FIG. 1; P-3 monohydrate crystal formcharacterized by a powder X-ray diffraction pattern as shown in FIG. 9;P-3 dihydrate crystal form characterized by a powder X-ray diffractionpattern as shown in FIG. 8; P-3 ethanolate crystal form characterized bya powder X-ray diffraction pattern as shown in FIG. 11; P-3 anhydrouscrystal form characterized by a powder X-ray diffraction pattern asshown in FIG. 10; P-2 anhydrous crystal form characterized by a powderX-ray diffraction pattern as shown in FIG. 2; P-4 crystal formcharacterized by a powder X-ray diffraction pattern as shown in FIG. 19;P-6 crystal form characterized by a powder X-ray diffraction pattern asshown in FIG. 20; P-7 crystal form characterized by a powder X-raydiffraction pattern as shown in FIG. 21; P-8 crystal form characterizedby a powder X-ray diffraction pattern as shown in FIG. 22; andcombinations thereof.
 15. The composition of claim 14, wherein thecomposition comprises P-2 hydrate crystal form characterized by a powderX-ray diffraction pattern as shown in FIG.
 1. 16. The composition ofclaim 14, wherein the composition comprises P-3 monohydrate crystal formcharacterized by a powder X-ray diffraction pattern as shown in FIG. 9.17. The composition of claim 14, wherein the composition comprises P-3dihydrate crystal form characterized by a powder X-ray diffractionpattern as shown in FIG.
 8. 18. The composition of claim 14, wherein thecomposition comprises P-3 ethanolate crystal form characterized by apowder X-ray diffraction pattern as shown in FIG.
 11. 19. Thecomposition of claim 14, wherein the composition comprises P-3 anhydrouscrystal form characterized by a powder X-ray diffraction pattern asshown in FIG.
 10. 20. The composition of claim 14, wherein thecomposition comprises P-2 anhydrous crystal form characterized by apowder X-ray diffraction pattern as shown in FIG.
 2. 21. The compositionof claim 14, wherein the composition comprises P-4 crystal formcharacterized by a powder X-ray diffraction pattern as shown in FIG. 19.22. The composition of claim 14, wherein the composition comprises P-6crystal form characterized by a powder X-ray diffraction pattern asshown in FIG.
 20. 23. The composition of claim 14, wherein thecomposition comprises P-7 crystal form characterized by a powder X-raydiffraction pattern as shown in FIG.
 21. 24. The composition of claim14, wherein the composition comprises P-8 crystal form characterized bya powder X-ray diffraction pattern as shown in FIG.
 22. 25. Thecomposition of claim 14, additionally comprising one or morepharmaceutically acceptable carriers.
 26. The composition of claim 14,wherein the at least one polymorph of MDT-637 is dispersed in at leastone pharmaceutical excipient to provide a solid composite.
 27. A methodof treating a subject having a disease resulting from infection byParamyxovirinae or Pneumovirinae, comprising administering to thesubject a compound in an amount effective to treat the subject whereinthe compound comprises at least one polymorph of MDT-637 selected fromthe group consisting of: P-2 hydrate crystal form having characteristicpowder X-ray diffraction peaks at 2θ values of 4.83°, 8.42°, 9.61°,11.83°, 14.60°, 16.94°, 19.25°, 20.46°, 24.09°, 24.85°, 27.76° and28.56°; P-3 monohydrate crystal form having characteristic powder X-raydiffraction peaks at 2θ values of 8.25°, 9.55°, 11.66°, 14.30°, 18.03°,19.70°, 20.29°, 24.55°, 25.29°, 26.31° and 27.26°; P-3 dihydrate crystalform having characteristic powder X-ray diffraction peaks at 2θ valuesof 7.03°, 8.16°, double peak at 9.47° and 9.75°, 11.60°, 14.24°, 17.95°,19.64°, 20.26°, 24.52°, 26.24° and 27.25°; P-3 ethanolate crystal formhaving characteristic powder X-ray diffraction peaks at 2θ values of8.25°, 9.54°, 9.85°, 11.69°, 14.32°, 15.99°, 18.06°, 19.73°, 20.33°,24.54°, and 26.27°; P-3 anhydrous crystal form having characteristicpowder X-ray diffraction peaks at 2θ values of 8.27°, 9.54°, 11.68°,14.29°, 18.07°, 19.79°, 20.30°, 24.54°, 25.27°, 26.36°, and 27.25°; P-2anhydrous crystal form having characteristic powder X-ray diffractionpeaks at 2θ values of 8.58°, 10.64°, 12.58°, 14.51°, 15.89°, 17.20°,21.06°, 21.41°, 22.37°, 25.43° and 27.62°; P-4 crystal form havingcharacteristic powder X-ray diffraction peaks at 2θ values of 4.31°,7.99°, 9.37°, 11.02°, 13.04°, 13.43°, 14.18°, 16.13°, 16.70°, 17.08°,17.42° and 17.92°; P-6 crystal form having characteristic powder X-raydiffraction peaks at 2θ values of double peak at 3.43° and 3.89°, 6.89°,double peak at 7.87° and 8.25°, 10.88°, 13.06°, 13.81°, 16.12°, 17.38°and 18.51°; P-7 crystal form having characteristic powder X-raydiffraction peaks at 2θ values of 5.18°, 6.59°, 7.70°, double peak at10.02° and 10.62°, double peak at 12.18° and 12.50°, 15.16°, double peakat 15.88° and 16.4°, double peak at 17.20° and 17.42°, and double peakat 18.07° and 18.25°; P-8 crystal form having characteristic powderX-ray diffraction peaks at 2θ values of 4.15°, 7.85°, 9.33°, 14.29°,triple peak at 15.84°, 16.68° and 17.14°, 18.53°, 20.27°, 23.90°, 24.80°and 27.39°; and combinations thereof.
 28. The method of claim 27,wherein the method of treating the disease comprises alleviating andpreventing symptoms associated with respiratory syncytial virus (RSV),wherein the symptoms comprise rhinitis, otitis media, pneumonia andbronchiolitis.
 29. The method of claim 27, wherein the compound isadministered to the subject via inhalation.
 30. A method of treating asubject having a disease resulting from infection by Paramyxovirinae orPneumovirinae, comprising administering to the subject a compound in anamount effective to treat the subject wherein the compound comprises atleast one polymorph of MDT-637 selected from the group consisting of:P-2 hydrate crystal form characterized by a powder X-ray diffractionpattern as shown in FIG. 1; P-3 monohydrate crystal form characterizedby a powder X-ray diffraction pattern as shown in FIG. 9; P-3 dihydratecrystal form characterized by a powder X-ray diffraction pattern asshown in FIG. 8; P-3 ethanolate crystal form characterized by a powderX-ray diffraction pattern as shown in FIG. 11; P-3 anhydrous crystalform characterized by a powder X-ray diffraction pattern as shown inFIG. 10; P-2 anhydrous crystal form characterized by a powder X-raydiffraction pattern as shown in FIG. 2; P-4 crystal form characterizedby a powder X-ray diffraction pattern as shown in FIG. 19; P-6 crystalform characterized by a powder X-ray diffraction pattern as shown inFIG. 20; P-7 crystal form characterized by a powder X-ray diffractionpattern as shown in FIG. 21; P-8 crystal form characterized by a powderX-ray diffraction pattern as shown in FIG. 22; and combinations thereof.31. The method of claim 30, wherein the method of treating the diseasecomprises alleviating and preventing symptoms associated withrespiratory syncytial virus (RSV), wherein the symptoms compriserhinitis, otitis media, pneumonia and bronchiolitis.
 32. The method ofclaim 30, wherein the compound is administered to the subject viainhalation.